小麦胚乳特异性启动子的克隆及高分子量谷蛋白14亚基基因的原核表达
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摘要
由多个亚基组成的高分子量（HMW）谷蛋白是小麦种子中一类重要的贮藏蛋白，
占种子总蛋白的量的 10%以上。大量研究表明，小麦高分子量谷蛋白亚基（HMW-GS）
组成与小麦的加工品质密切相关[1]。因此，利用基因工程将优质亚基导入小麦栽培品种
以改良其品质是当前小麦品质改良遗传育种的重要课题。因此得到一个小麦优质亚基基
因和能驱动此基因在小麦种子中专一且高效表达的启动子，是实现这一目标的前提。
小麦 HMW-GS 是一类种子贮藏蛋白，它的表达具有很高的胚乳专一性。因此，我们
利用 PCR 技术克隆了 HMW-GS 基因的启动子。通过对启动子序列的同源性及聚类分析，
结果表明：本实验所克隆的启动子为小麦 HMW-GS 1Dx2 启动子，长度约为 1050bp，
GenBank 登陆号为:AJ577815。将此片段与β-葡萄糖苷酸酶(Gus)基因编码序列融合，再
用基因枪法将其转入小麦种子及叶片中。组织化学检测表明，GUS 基因只在种子得到表
达，并比 CaMV 35S 启动子驱动的 GUS 在种子中表达量更高，因此从功能上证明了此
HMW-GS 启动子片段能指导外源基因在小麦种子中高效且专一的表达。
小麦种子特异性启动子的分离，为利用小麦种子作为反应器来生产特殊蛋白奠定基
础，也是基因工程技术改良小麦品质的前提。在对小麦进行品质改良分子育种时，除拥
有自主知识产权的胚乳专一性启动子外，还需要拥有一些与品质呈正相关的优良基因。
研究表明：小麦的加工品质与小麦种子中麦谷蛋白和醇溶蛋白的含量和组成关系极为密
切，在不同高分子量麦谷蛋白亚基对加工品质的贡献的研究中发现 5+10 亚基为优质亚
基，而 2+12 亚基为劣质亚基。然而我国的一些优质面包小麦品种如小偃 6 号（亚基组成
为 1，14+15，2+12）等缺乏 5+10 亚基，而具有认为是劣质的 2+12 亚基。分析结果表明
导致其优质的原因在于其所具有的 14+15 亚基，14+15 亚基可能具有 5+10 亚基的功能。
因此，研究 14 亚基基因的功能对改良小麦品质具有重要意义。本研究在大量分析
HMW-GS14 基因序列及总结以往实验结果的基础上，利用改进的 PCR 缓冲体系及一对
高 Tm 值的引物，通过二段法成功的克隆了 HMW-GS14 基因。将 HMW-GS14 基因插入
到原核表达载体 pET30a(+)质粒上，并使重组质粒在大肠杆菌中得到表达，为验证
HMW-GS14 基因的功能奠定基础，也为利用基因工程改良小麦作准备。
High-molecular-weight glutenins which constitute of many subunits is a kind of
important storage protein and occupy 10% of whole protein in seed. Plenty of studies showed
that constitution of high-molecular-weight glutenin subunit (HMW-GS)play a signification role
on Triticeae processing quality. It is an important subject that transfer gene into cultivate
Triticeae to improve quality utilizing gene engineering. So receiving a quality gene and
promoter which can drive it specially expression in embryo is a precondition to quality
breeding.
Triticeae high-molecular-weight glutenins is a important seed storage protein and
specially expression in embryo. We cloned a promoter utilizing PCR technology. Sequencing,
homology and alignment analysis showed that the cloned product is HMW-GS 1Dx2’s
promoter, the length is 1041 base pairs and the number of Genbank is AJ577815. Linked it
with Gus gene and transferred it into Triticeae seed and slide with gene gun, only observed
instant expression product in Triticeae seed, which certificated that HMW-GS promoter can
direct reporting gene especially expression in Triticeae seed.
Isolation specially of Triticeae seed settle foundation to use plant as vector to produce
protein and also is precondition to improve Triticeae quality utilizing gene engineering
technology. During improve Triticeae quality must have some quality genes and specially
promoters. The content and constitutes of glutenins and prolamin in Triticeae seed with
processing quality have closed relation. During the analysis contribute of different HMW-GS
to processing quality find that 1Dx5+1Dy10 is quality subunits but 1Dx2+1Dy12 is bad
subunits. Some quality bread Triticeae such as xiaoyan 6 (1Bx14+1By15 and
1Dx2+1Dy12)have no 1Dx5+1Dy10 but have 1Dx2+1Dy12 in our country. The analysis
showed that 1Bx14+1By15 have the same function with 1Dx5+1Dy10, so research
1Bx14+1By15 have signification to quality breeding. We abundance analysis HMW-GS14
gene sequence, summarize anciently experimental result and succeed received HMW-GS14
gene making use of ameliorative PCR buffer system and a pair of high melting temperature(Tm)
primers. Insert it into prokaryotic expression vector pET30a(+), gained recombinant plasmid
pET30a-HMW and transformed pET 30a -HMW into E.coli and were expressed intention
protein product which establish foundation to function identification HMW-GS14 and improve
Triticeae utilizing gene engineering.
占种子总蛋白的量的 10%以上。大量研究表明，小麦高分子量谷蛋白亚基（HMW-GS）
组成与小麦的加工品质密切相关[1]。因此，利用基因工程将优质亚基导入小麦栽培品种
以改良其品质是当前小麦品质改良遗传育种的重要课题。因此得到一个小麦优质亚基基
因和能驱动此基因在小麦种子中专一且高效表达的启动子，是实现这一目标的前提。
小麦 HMW-GS 是一类种子贮藏蛋白，它的表达具有很高的胚乳专一性。因此，我们
利用 PCR 技术克隆了 HMW-GS 基因的启动子。通过对启动子序列的同源性及聚类分析，
结果表明：本实验所克隆的启动子为小麦 HMW-GS 1Dx2 启动子，长度约为 1050bp，
GenBank 登陆号为:AJ577815。将此片段与β-葡萄糖苷酸酶(Gus)基因编码序列融合，再
用基因枪法将其转入小麦种子及叶片中。组织化学检测表明，GUS 基因只在种子得到表
达，并比 CaMV 35S 启动子驱动的 GUS 在种子中表达量更高，因此从功能上证明了此
HMW-GS 启动子片段能指导外源基因在小麦种子中高效且专一的表达。
小麦种子特异性启动子的分离，为利用小麦种子作为反应器来生产特殊蛋白奠定基
础，也是基因工程技术改良小麦品质的前提。在对小麦进行品质改良分子育种时，除拥
有自主知识产权的胚乳专一性启动子外，还需要拥有一些与品质呈正相关的优良基因。
研究表明：小麦的加工品质与小麦种子中麦谷蛋白和醇溶蛋白的含量和组成关系极为密
切，在不同高分子量麦谷蛋白亚基对加工品质的贡献的研究中发现 5+10 亚基为优质亚
基，而 2+12 亚基为劣质亚基。然而我国的一些优质面包小麦品种如小偃 6 号（亚基组成
为 1，14+15，2+12）等缺乏 5+10 亚基，而具有认为是劣质的 2+12 亚基。分析结果表明
导致其优质的原因在于其所具有的 14+15 亚基，14+15 亚基可能具有 5+10 亚基的功能。
因此，研究 14 亚基基因的功能对改良小麦品质具有重要意义。本研究在大量分析
HMW-GS14 基因序列及总结以往实验结果的基础上，利用改进的 PCR 缓冲体系及一对
高 Tm 值的引物，通过二段法成功的克隆了 HMW-GS14 基因。将 HMW-GS14 基因插入
到原核表达载体 pET30a(+)质粒上，并使重组质粒在大肠杆菌中得到表达，为验证
HMW-GS14 基因的功能奠定基础，也为利用基因工程改良小麦作准备。
High-molecular-weight glutenins which constitute of many subunits is a kind of
important storage protein and occupy 10% of whole protein in seed. Plenty of studies showed
that constitution of high-molecular-weight glutenin subunit (HMW-GS)play a signification role
on Triticeae processing quality. It is an important subject that transfer gene into cultivate
Triticeae to improve quality utilizing gene engineering. So receiving a quality gene and
promoter which can drive it specially expression in embryo is a precondition to quality
breeding.
Triticeae high-molecular-weight glutenins is a important seed storage protein and
specially expression in embryo. We cloned a promoter utilizing PCR technology. Sequencing,
homology and alignment analysis showed that the cloned product is HMW-GS 1Dx2’s
promoter, the length is 1041 base pairs and the number of Genbank is AJ577815. Linked it
with Gus gene and transferred it into Triticeae seed and slide with gene gun, only observed
instant expression product in Triticeae seed, which certificated that HMW-GS promoter can
direct reporting gene especially expression in Triticeae seed.
Isolation specially of Triticeae seed settle foundation to use plant as vector to produce
protein and also is precondition to improve Triticeae quality utilizing gene engineering
technology. During improve Triticeae quality must have some quality genes and specially
promoters. The content and constitutes of glutenins and prolamin in Triticeae seed with
processing quality have closed relation. During the analysis contribute of different HMW-GS
to processing quality find that 1Dx5+1Dy10 is quality subunits but 1Dx2+1Dy12 is bad
subunits. Some quality bread Triticeae such as xiaoyan 6 (1Bx14+1By15 and
1Dx2+1Dy12)have no 1Dx5+1Dy10 but have 1Dx2+1Dy12 in our country. The analysis
showed that 1Bx14+1By15 have the same function with 1Dx5+1Dy10, so research
1Bx14+1By15 have signification to quality breeding. We abundance analysis HMW-GS14
gene sequence, summarize anciently experimental result and succeed received HMW-GS14
gene making use of ameliorative PCR buffer system and a pair of high melting temperature(Tm)
primers. Insert it into prokaryotic expression vector pET30a(+), gained recombinant plasmid
pET30a-HMW and transformed pET 30a -HMW into E.coli and were expressed intention
protein product which establish foundation to function identification HMW-GS14 and improve
Triticeae utilizing gene engineering.
引文
1. 苟吉庆. 植物顶端组织高效诱导型启动子的创建与表达调控研究，[博士论文]
2001
2. 刘良式等. 植物分子遗传学. 1997，492
3. 刘良式等. 植物分子遗传学. 1997，210
4. 刘昱辉，王志幸，贾仕荣. 拟南芥 profiling 启动子 5’端缺失对维管束特异表达
的影响. 科学通报, 2001，5:835-828
5. 吴标，潘瑞琴，芦睿，田颖川. 竹节花黄斑驳病毒启动子的缺失分析及功能. 微
生物学报, 1999，39 (1):15- 21
6. 蒋浩，秦红敏，虞红梅. 笋瓜韧皮部蛋白基因启动子的克隆及功能初探. 农业生
物技术学报，1999，7 (1): 63-68
7. 黄伟达，姜学军，黄皓，贺斌等. 小麦高分子量谷蛋白基因转录调控相关的顺反
式相互作用的初步研究. 植物学报，1995，37:425-431
8. 李胜国，刘玉乐，康良仪，田波. 烟草花药特异性启动子的克隆、活性测定及雄
性不育基因和恢复基因的构建. 农业生物技术学报，1993，3 (3): 25-32
9. 杨洪全，卫志明，许智宏. Rch10 启动子引导 iaaL 基因在转荃因烟草中的表达导
致花器官发育的变异. 植物学报，1996，38 (7):541 – 547
10. 范惠琴，姚泉洪，黄晓敏，彭日荷. 矮牵牛查尔酮黄烷酮异构酶（chiA）基因启
动子的克隆和测序. 上海农业学报, 1998，14(3):14-16
11. 叶梁，李极，宋艳茹. 双价、三价种子特异性表达载体的构建及表达 PHB 合成
相关基因的转基因油菜的获得. 科学通报，2000，45(5):516
12. 赵友梅，王淑俭. 高分子量麦古蛋白亚基的 SDS-PAGE 图谱在小麦品质研究中
的应用. 作物学报，1990，16(3):208-218
13. 王瑞等. 一些优质小麦及其杂种后代高分子量谷蛋白亚基组成与面包品质之关
系. 西北农业学报, 1995，4:25－30
14. 刘香利. 小麦醇溶蛋白和高分子量麦谷蛋白亚基组成与品质关系研究，[硕士论
文] ，1999
15. 徐虹. 中间偃麦草麦谷蛋白亚基的鉴定、基因克隆与分子特征分析, [博士论文]，
2003
16. 王关林，方宏筠主编. 植物基因工程. 科学出版社，北京 2002，143-172
17. 解瑞立，万永芳，张慧，王道文. 多倍体山羊草物种中高分子量麦谷蛋白亚基组
成与其编码基因的分子克隆. 科学通报，2000，45(17):1867-1874
18. 张晓东，李冬梅，徐文英等. 利用基因枪将 HMW 谷蛋白亚基基因与除草剂 Basta
抗性基因导入小麦不同外植体获得转基因植株。遗传 HEREDITAS（Beijin）（增
56 小麦胚乳特异性启动子的克隆及高分子量谷蛋白 14 亚基基因的原核表达
刊），1998，20:3－8
19. 陈梁鸿，王新望，张晓东等. 小麦编码高分子量谷蛋白亚基基因的转化. 作物学
报，1999，25(4):437－440
20. 唐凤兰等. 利用花粉管通道法向小麦中导入麦谷蛋白高分子量优质亚基基因的
研究初报. 黑龙江农业科学，2002，3:1－2
21. 赵虹. 小麦基因枪转化体系的建立和抗除草剂基因的导入, [硕士论文]， 2001
22. 邓志勇. 高分子量麦谷蛋白 14 和 15 亚基的纯化及部分生化特性的研究, [硕士论
文] ，1999
23. 范三红. 小麦高分子量麦谷蛋白 14 和 15 亚基的研究, [硕士论文]， 2001
24. Donal, R,G. kand cashmore, A.R. RG Donald and AR Cashmore. Mutation of
either G box or I box sequences profoundly affects expression from the Arabidopsis
rbcS-lA promoter. EMBO J (1990) 9: 1717-1726
25. P Puente, N Wei, and XW Deng. Combinatorial interplay of promoter elements
constitutes the minimal determinants for light and developmental control of gene
expression in Arabidopsis. EMBO J (I996) 15: 3732-3743
26. Hawk-Bin Kwon, Soo-Chul Park. Identification Of a Light Responsive Region of
the Nuclear Gene Encoding the B Subunit of Chloroplast Glyceraldehyde
3-Phosphate Dehdrogenase from Arahidopsis thaliana. Plant Physiol(1994)
105:357-367
27. Slogan JS, Schwartz BW, Becker WM: Promoter analysis of a light of a
light-regulated gene encoding Hydroxyyruvate reductase, an enzyme of the
photorespi -ratory glycolate pathway. Plant J(1993)3:867-874
28. Steven G. Daniel, Wayne M. Becker. Trangenic analysis the 5'-and 3'-flanking
regions of the NADH-dependent hydroxypyuvate redutctase gene from Cucumis
sativus L. Plant-molecuar-biology (1995) 28:821-836.
29. Lagrange,T.,B. Axeloss,M. Structure and expression of the nuclear gene coding for
the chloroplast ribosmal protein L2: Developmental regulation of a house-keeping
gene by altemative promoters. MoI. Cell. Biol (1993) 13:2614-2622
30. Harrak, H. Largrange, T. The expression of nuclear genes encoding plastid ribosomal
precedes the experssion of chloroplast genes during early phases of chloroplast
development. Plant physiol(1995)108:685-692
31. Zhou,D.-X. Li,Y-F. Sequence specific interaction between SIF, a spinach nuclear
factor,and a negative cis-element conserved in plastid-related genes. J. Biol. Chem
(1992) 267:23515-23519
32. Thiery Lagrange, Syiviane Gauvin. S2F, a leaf-Specic trans-Acting Factor, Binds to a
参考文献 57
Novel cis-Acting Element and Differentially Activates the RPL21 Gene. The plant
Cell(1997) 9:1469-1479
33. Ke11er,B. Baumgartner, C. Vascualr-specific expression of the bean GRP1.8 gene is
negatively regulated. Plant cell (1991) 10:1051-1061
34. Hatton,D.Sablowski,R.,Yung M H ,et al. Two classes of cis sequences contribute to
tissue -specific expression of a PAL2 promoterr in transgenic tobacco.The plant
J(1995)7(6):859-876
35. Yin Y，Chen L，Beachy R.1997.Promoter elements required for phloem-specific gene
expression from the RTBV promoter in rice.Plant J.12(5):1179-88
36. Graham MW，Craig S，Waterhouse PM.1997.Expression patterns of vascular -specific
promoters Ro1C and Sh in transgenic potatoes and their use in engineering
PLRV-resistant plants.Plant Mol Biol.33(4):729-35
37. Koltunow,AN.,et al .Different Temporal and Spatial Gene Expression Patterns Occur
during Anther Development .Plant Cell(1990)2:1201-1224.
38. Schrauwen JAM，de Groot PFM，van Herpen MMA，van der Lee T，Reynen WH，
Weterings KAP，Wullems GJ.1990.Stage-related expression of mRNAs during pollen
development in lily and tobacco.Planta，182:298-304
39. Hanson DD ， Hamilton DA ， Travis JL ， Bashe DM ， Nascarenhas JP.
1989.Characterization of a poolen-specific cDNA clone from Zea mays and its
expression. Plant Cell,1:173-179.
40. Zou J-T, Zhan X-Y, Wu H-M, Wang H, Chueng AY. 1994.Characterization of a rice
pollen-specific gene and its expression.American Journal of Botany，81:552 –561
41. Pe ME, Frova C，Colombo L，Bineli G，Mastroianni N，Tarchini R，Visioli
G.1994.Molecular cloning of genes expressed in pollen of Sorghum bicolor.
Maydica，39:107-113
42. Batanero E ， Villaba M ， Lopez-Otin C ， Rodriguez R.1994.Isolation and
characterization of an olive allergen-like protein from lilac pollen.Sequence analysis
of three cDNA encoding protein isoforms.Eur J Biochem，221:187-193
43. Lombardero M.，Barbas JA，Moscoso del Prado J，Carreira J，1994.cDNA sequence
analysis of the main olive allergen.Olee I.Clin Exp Allergy，24:765-770
44. Twell D，Yamaguchi J，McCormick S，1990.Pollen-specific gene expression in
transgenic plants:Coordinate regulation of two different tomato gene promoters
during microsporrgenesis.Development，109:705-713
45. Green PJ.Yong MH Cuozzzo M ， kano-Murakami Y ， Silverstein P. ， Chua
N-H.1988.Binding site requirements for pea nuclear protein factor GT- correlate with
58 小麦胚乳特异性启动子的克隆及高分子量谷蛋白 14 亚基基因的原核表达
sequences required for ligft-dependent transcriptional activation of the rbcS-3A gene.
EMBO J, 7:4035-4044
46. Kuhlemeier C ， Cuozzo !M ， Green PJ ， Goyvaerts E ， Ward K ， Chua
N-H .1988.Localition and conditional redundancy of regulatory elements in rbcS-3A，
apea gene encoding thr small subunit of ribulose-bisphospgate carboxylase.Proc Natl
Acad Sci USA，85:4662-4666
47. Twell D，Yamaguchi J，Wing R，McCormick S，1991.Promotei analysis of three
genes that are coordinately expresse during pollen development reveals pollen
specific enhances sequences and shared regulatory elements.Genes Devel.，5:496-507
48. Eadl Y，Curie C，McCormick S.Pollen specificity elements reside in 30bp of the
proximal promoters of two polloen expressed genes.Plant Cell，7:373-384
49. N.Bate&D.TWell.1998.Functional architecture of a late pollen
promoter:pollen-specific transcription is developmentaaly regulated by multiple:tage
specific and dependent activator elements .Plant Molecular Biology，37:859-869
50. De Pater,S.,Pham,K. J. A 22-by Fragment of the Pea Lectin Promoter Containing
Essential TGAC-like Motifs Confers Seed-Specific Gene Expression .Plant cell
(1993)5:877-886.
51. Bustos,M.M.,Vegum,D. Positive and negative cis-acting DNA domains are required
for spatial and temporal regulation of gene expression by a seed storage protein
promoter .EMBOJ(1991)10:1469-1479
52. Marzable,P,Busk,P,K Ludevid,M.D .and Torrent,M. The bifactorial endosperm box
of r-zein gene:characterization and functional of thePb3 and GZM scis-acting
elements. Plant J(1998).16:41-52
53. Vicen-Carbajosa,J.,Onate,L.,Lara,P.,Diaz,L.and Carbonero,P.,Barly BLZ1:A bZIP
transcriptional activator that interact with endosperm-specific gene promoters. Plant
J(l998)13:629-640
54. Takaiwa,F.,Yamanouchi,U.,Yoshihara,T.,Washida,H.,Tanabe,F.,Kato,A.
Characterization of common cis-regulatory elements responsible for the
endosperm-specific expression of members of the rice glutelin multigene family.
Plant MOL.Biol(1996)3:1207-1221
55. Washida,H.,Wu,C.-Y,Yamanouchi,U.,akihanma,T.,Harada,K.and Takaiwa,F.
Identification of cis-regulatory elements required for endosperm expression of the
storage protein glutelin gene Glub-1. Plant MOL.Biol(1999)40,1-12
56. Zheng,Z,Kawagoe,Y,xiao,S,Li,Okita,T,Hau,TL,Lin,A.and Murai,.N. 5’distal and
proximal cis-acting regulatory elements are riquired for developmental control arice
参考文献 59
seed storagegene. Plant J(1993)4:357-366.
57. Yoshihra,Twashida,H.and Takaiwa,F.A 45-by proximal regioncontaining AACA and
GCN4 motif is suffient to confer endosperm-specific expression of the rice storage
protein glutein gene Glu-3. FEBS Lett(1996)383:213-218
58. Albani,D.,Hammond-Kosack,M.C.U., The wheat transcriptional activator SPA:a
seed-specific bZJP protein that recognizes the GCN4-like motif in the bifactoral
endosperm box of prolamin genes .Plant Cell(1997) 9:171-184
59. Mena,M.,Vicente-Carbajosa,J.,Schmidt,R.J. and Caarbonero,P. An
endosperm-specific DOF protein from barly,highly conserved in wheat ,binds to and
activies transciption from the prolamin -box of a native B- hordein promoter in barly
edospeerm box. Plant J(1998)16:53-62
60. Suzki,A.,Wu,C.-Y Washida,H.and Takaiwa,F. Rice NM protein OSMYB5
specifically binds to the AACA motif conserved among promoters of gene for storage
protein gllutelin. Plant cell Physiol(1998)39:555-559.
61. Schmidt,R.,Katagiri,M. Opaque-2 is a transcriptional activator that recognizes a
specific targert site in 22kD zein genes.Plant cell(1992) 4:689-700.
62. Wu,C.-YSuzuki,A.,Washida,H.and Takaiwa,F.The GCN4 motif in a rice glutilin gene
is essential for endosperm-specific gene expression and is actived by Opaque-2 in
transgenic rice plants. Plant J(1998)14,673-683
63. Chuan-Yin Wu,Haruhiko Washida.Quantitatve nature of the Prolamin-box,ACGT
and AACA motifs in a rice glutelin gene promoter:Minimal cis –element riquiremints
for endosperm-specific gene.The plant Journal (2000)23:415-421
64. Washida,H.,Wu,C.-Y,Yamanouchi,U.,akihanma,T.,Harada,K.and Takaiwa,F.
Identification of cis-regulatory elements required for endosperm expression of the
storage protein glutelin gene Glub-1. Plant MOL.Biol(1999)40,1-12
65. Robert L S,Thompson R D,Flavell R B.Tissue-specific expression of a wheat high
molecular weight glutenin gene in transgenic tobacco.Plant Cell,1989,1:569-578
66. Bate，N.，Spurr，C.，Forter，G.D.and Twell，D.1996. Maturation 一 specific
translational enhancement mediated by the 5，-UTR of a late pollen transcript. Plant
J.10:613-623
67. D. Hatton，R.Sablowski，M-H Yung，G.Smith，W. Schuch and M. Bevan.1995.Two
classes of cis sequences contribute to tissue-specific expression of a PAL2 promoter
in transgenic tobacco.The Ptant J.，7(6):859-876
68. Bronstein I, Martin CS, Fortin JJ, et al. Chemiluminescence: sensitive detection
technology for reporter gene assays［J］. Clin Chem,1996,42(9): 1542.
60 小麦胚乳特异性启动子的克隆及高分子量谷蛋白 14 亚基基因的原核表达
69. Koziel M. G.，BelandG.L.，Bowman C.，N.B.Carozzi，R.Crenshaw, et al，1993.Field
performance of elitee transgenic maize plants expressing an insecticidal protein
derived from Bacillus thuringiensis. Bio/Technology，11:194-200
70. Oxtoby，E.，Hughes，M.A.，1990.Engineering herbicide tolerance into crops.Trends
Biotechnol.8:61-65
71. Edwards，J.W.，Waiker，E.L.，Coruzzi，G.M.1990.Cell-specific expression in
transgenic plants reveals non-overlapping roles for chloroplast and cytosolic
glutamine synthetase .Proc.Natl.Acad.Sci.USA，87:3459-3463
72. Mol，J.，Stuitje，A.，van der Krpl，A.，Jorgensen，R.1989. Saying it with
genes:Molecular flower breeding.Trebds Biotechnol.7:148-153
73. Tunen A, Mur L, Mol JNM. 1990. Pollen- and anther-specific chia promoters from
petunia:Tandem promoters regulation of the chia gene. The Plant Cell, 2:393-401
74. Sato ， T. ， Theologis ， A. 1989.Cloning the mRNA encoding
1-ammminocyclopropane-1-carboxylate synthase, the key enzyme for ethylene
biosynthesis in plants.Proc.Natl.Acad.Sci .USA. 86:6621-662
75. Edwards，J.W.，Waiker，E.L.，Coruzzi，G.M.1990.Cell-specific expression in
transgenic plants reveals non-overlapping roles for chloroplast and cytosolic
glutamine synthetase .Proc.Natl.Acad.Sci.USA，87:3459-3463
76. Vandekerckhove，i.，Damme，i.F.，Lijsebettens，M.，Botterman，i.，De-Block，
M.，1989.Enkephalins produced in trangenic plants using modified 2S seed storage
proteins.Biotechnoloav.7:929-932
77. Firek S.，Draper J，Owen MRL.Gandecha A.，Cockburn W.，Whitelam GC. Secretion
of a functional single 一 chian Fv protein in transgenic tobacco plants and cell
suspension.1993
78. Ma JK-C，Hiatt A.，Hein M.，Vine ND.，Wang F.，Stab ila P.，van Dolleweerd C.，
Mostov K.，Lehner T.，1995.Generation and assembly of secretory antibodies in
plants.Science，268:716-719
79. Mason HS，Lam DM-K，Arntzen CJ. 1992.Expression of hepatitis B surface antigen
in transgenic plants.Proc Natl.Acad.Sci.USA, 89:11745-11749
80. Haq TA，Mason H. S.，Clements JD.，Arntzen CJ.，1995.Oral immunization with a
recombinant bacterial antigen produced in transgenic plants. Science, 268:714-716
81. Matsumoto S，I kura K，Ueda M，Sasaki R.1995.Characterization of a human
glycoprotein(erythropoietin)produced in cultutred tobacco cell，.Plant Mol.Biol.
27:1163-1172.
82. Ottoboni LMM，Lei te A，Yunes JA，Targon MLPN，de Souza Filho GA, Arruda P.，
参考文献 61
1993. Sequence analysis of 22kDa-like a-coixin and their comparison with
homologous zein and kafirin genes reveals highly conserved protein structure and
regulatory elements.Plant Mol Biol.，21:765-778
83. Adilson L，Edson L.K，Marc i o J.da S，Augusto D.L.，Rodrigo M. P .S.，Eric D .B.，
Hamza F.E-D.，Paulo A.，2000.Expression of correctly processed human growth
hormone in seeds of transgenic tobacco plants.Molecular Breeding，6:47-53
84. Wahlund KG, Gustavsson M, MacRitchie F, Nylander T,Wannerberger L(1996) Size
characterization of wheat proteins, particularly the gluteins, byasymmetrical flow
field-flow fractionation. J. Cereal Science 23:113-119
85. Stevenson SG, Prestom KR(1996) Flow Field-Flow Fractionation of Wheat Protein
Cereal Sci. 23:121-131
86. Wrihley CW(1996) Giant proteins with flour power. Nature 381:738-739
87. Seilmeier W, Belitz HD, Wieser H. Separation and quantitative determination of
high-molecular-weight subunit of glutenin from different wheat varieties and genetic
variants of the variety Sicco. Zeitschrift fur Lebensmittel Untersuchung
und-Forschung ，1991，192: 124-129
88. Weegles Pl, Hamer RJ, Schofield JD(1995) RP-HPLC and Capillary Electrophoresis
of Subunits from Glutein Isolated by SDS and Osborne Fraction. J Cereal Sci.
22:211-224
89. Vasil V,Castillo A M,Fromm M E and Vasil I K. Herbicide resistant fertile
transgenetic wheat plants obtained by mircoprojectile bombardment of regenerable
embyogenic callus, Bio/technology. 1992,10:667-674
90. Fredy Altpter,Vimla vasil,Vibbas Srivastava and Indra K Vasil. Intergration and
expression of the High-Moleclure-weight lutenin subunits 1AxI gene into wheat
Nature Biotechnology 1996, 14:1155-1159
91. Ann E Bkchl and Olin d Anderson . Expression of a Novel High-Molecular -weight
Glutenin subunit gene in transgenic wheat. Nadture Biotechnology 1996,14:875-879
92. Rooke L,Barro F,Tatham A S,Fido R,Steel S,Bekes F,Cras P,Martin A,Lazzeri P
A,Shewry P R, Barccelo P. 1999,Altered functional properties of tritordeum by
transformation with HMW glutein subunit genes.Theoretical And Applied Genetics.
99:851-859
93. Payne PI, Holt LM, Lawrence GJ(1983) Detection of a novel high molecular weight
subunit of glutenin in some Japnese hexaploid wheats. J. Cereal Sci. 1:3-8
94. Payne PI, Corfield KG, Blackman JA(1981) correlation between the inheritance of
certain high-molecular-weight subunit of glutenin and bread-making quality in
62 小麦胚乳特异性启动子的克隆及高分子量谷蛋白 14 亚基基因的原核表达
progenies of six corsses of bread wheat. J. Sci. Food Agric. 32:51-60
95. Anderson,O.D.and F,C.Greene,The characterization and comparative analysis of high
molecular weight glutenin genes from genomes A and B of a hexaploid bread
wheat.Theor.Appl.Genet. 1989b ,77:689－700
96. Anderson,O.D.F.C.Greene.Nudeotide sequences of the two high－molecular weight
glutenin genes from the D － genome of hexaploid wheat,Triticum aestivum
L.CV.cheyene.Nucleic Acids Res. 1989a ,17:461－462
97. Anderson,O.D.Litts.J.C. Nucleic acid sequence and chromosome assignment of a
wheat storage protein gene.Nucleic Acids Res 1984,12:8129－8144
98. Forde,J.et al, The nucleotide sequence of a HMW glutenin subunit gene located on
chromosome 1A of wheat (Triticum aestivum L.). Nucl.Acids Res. 1985,13:6817－
6832
99. Halford ,NG The mucleotide and deduced amino acid sequences of an HMW glutenin
subunit gene from chromosome 1B of bread wheat (Triticum aestivum L.)and
comparison with those of genes from chromosomes 1A and 1D.Theor Appl Genet
1987,75:117－126
100. Shenry,P.R.et al 1992,High molecular weight subunits of wheat glutenin J.Cereal
Sci 15:105－120
101.Deng ZY, Zhao HX, Fan SH, Ji WQ, Guo AG, Xue XZ. Purification and
biochemical characterization of high-molecular-weight-glutenin subunits 14 and
15.Yi Chuan Xue Bao.; 2001,28(1):46-51. Chinese
102.Mackie AM, Sharp PJ, Lagudah ES. The Nucleotide and Derived Amino Acid
Sequence of a HMW Clutenin Gene from Triticum tauschii and Comparison with
those from the D Genome of Bread Wheat. Journal of Cereal Science,1996, 24:73-78
103.Bustos ad, Rubio P, Jouve N. Molecular characterization of the inactive allete of the
gene Glu-AI and the development of a set of AS-PCR markers for HMW glutenins of
wheat. Theor. Appl. Genet.2000,100: 1085-1094
104.Wan Y, Wang D, Shewry PR, Halfordb NG(2002) Isolation and characterization of
five novel high molecular weight subunit of glutein genes from Triticum timopheevi
and Aegilops cylindrical. Theor Appl Genet 104:828-839
105.Shewry P.R., Napier J.A., Tatham A.S.. Seed storage proteins: structures and
biosynethsis.The plant cell., 1995,7:945-956
106.Shewry P.R., and Halford N.G.. Cereal seed storage protein:structures, properties and
role in grain utilization. J. Exp. Bot. 2002, 53(370):947-958
107.Roberts L S., Thompson R.D., Flavell R.B. Tissue-specific expression of a wheat
参考文献 63
high-molecular-weight glutenin gene in transgenic tobacco. Plant cell.1989.
1:569-578
108.Ballesteros J, Alvarez B, Gimenez J, Ramirez C, Cabrera A, Martin A. Introgression
of 1Dx5+1Dy10 into Tritordeum. Theor Appl Genet. 2003. 106(4):644-8
109.Rooke L, Barro F, Tatham AS, Fido R, Steele S, Bekes F, Gras P, Martin A, Lazzeri
PA, Shewry PR, Barcelo P. Altered functional properties of tritordeum by
transgprmation with HMW glutein subunit genes. Theor Appl Genet. 1999a.
99:851-858
110.Sangtong V., Moran R., Chikwamba R., Wand K., Woodman-Clikeman, Long M.J.,
Lee M., Scott M.P. Expression and inheritance of the wheat Glu-1Dx5 gene in
transgenic maize. Theor Appk Genet. 2002. 105:937-945
111.Bartel D,Thompson R D, Rodthstein S. 1985 Synthesis of a wheat stroge protein
subunit in Eschcerichia coliusing Novel expression vector,Gene,35(1-2):157-167
112.Maruyama N,Ichisc K,Kasube T,Kishimoto T,Kawase S,Matsumura Y,Takeuchi
Y,Sawada T,Utsumi S.1998,Identification of major wheat allergens by of the
Escherichia coli expression system Eur J Biochem Aug 1,255 (3):739-745
113.Calili J M,Rousset M,Qualate C Q.Kasarda D D .1990.Use of recmbinant inbred
lines of wheat for study of associationsof high-molecular-weight glutenia subunit
alleles to quartitative traits I.Grain yield and quality predication tests. Theoretical
And Applied Genetics.79:321-330
114.Anderson OD, Greec FC,Halford NG,Shewry PR,Malpica-Romero J-M 1989
Nucleotide sequence of die two high-molecular-weight glutenin gene from thr
D-genome of a bexaploid bread wheat, Triticum aestivum L.ev. Cheyenne. Nucleic
Acid Research 17:461-462.
115.Coraggio I,Compagno C,Martegani E,Ranzi B M,Sale E,Alberghina L and Viotti A.
1986 Transcription and expression of zein sequence in yeast under natural plant or
yeast promters. EMBO Journal,5:459-465
116.Campagno C,Coraggio J,Ranzi B M,Alberghina A L,Viotti A and MarteganiE.
1987,Translation of regulation of expression of zein cloned in yeast under inducible
GAL promoter.Biochemical and biophysical research commeication,146:809-814
117.Neill J D,Litts J C,Andasan O D,Greene F C and Stiles J I.1987,Expression of a
wheatα-gliadin gene in Soccharomcyces carevisiae, Gene,55:303-317
118.Scheets K and Hedgeoth C. 1989.Expression of a wheat α-gliadin gene in
Saccharomcyces cerevisiae from a yeast ADH1 promoter. Journal of agricultural and
food chemistry.37:829-833
64 小麦胚乳特异性启动子的克隆及高分子量谷蛋白 14 亚基基因的原核表达
119.Bartel D,Altosaar I,Harbert N P,Barker R F and Thompaon R D.1986, molecular
analysis of γ-gladin gene families at yhe complex Gli-I locus of bread wheat
(T.aestivum L.).uTheoretical and applied Genetics.72:845-853
120.Miyamoto C,Smithe G E,Farred-Towt J,Chizzonile R,Summers M D and Ju G.
1985,Productin of human c-myc protein in insect cells infected with a baculovirus
expression vector. Molecular and cell biology .5:2860-2865
121.Emery V C aad Bisbop D H L,1987.The development of multiple expression vector
for high level synthesis of eukaryotic proteins:expression of LCMVN polyhedrin
protein in a recombinant baculovirus system. Protein Engineer,1:359-366
122.Matsuura Y,Posse R D,Overton H A,and Bioshop D H L.1987,Baculovirus
expression vectors:The requirements for high level expression of proteins ,including
glycoproteins, Journal of General Virology ,68:1233 –1250
123.Ng P K W and Busluk W.1989,Concerning the nomenclature of high moleculare
weight glutein subuntits. Journal of cereal science,9:53-60
124.Lee Y.K., Bekes F., Ciaffi M. The low molecular weight glutenin subunit protenins
of primitive wheats. Ⅳ. Functional properties of products from individual genes.
Theor Appl Genet 1999c. 98:149-155
125.Sissons M.J., Bekes F., Skerritt J.H. 1998. Isolation and Functionality testing of low
molecular glutein subunits Cereal Chem., 75(1):30-36
126.D’ovidio R., Anderson O.D., Madci S., Skeritt J. and Porecddu E. Construction of
novel wheat high-Mr glutein subunit gene variability:modification of the repetitive
domain and exression in E.coli. Journal of cereal science. 1997b.25:1-8
127.Anderson O.D., Kuhl J C., Tam A. Construction and expression of a synthetic wheat
storage protein gene.Gene. 1996. 174:51-58
128.Blechl A E,Anderson O D. Expression of a novel high-molecular-weight glutenin
subunit gene in transgenic wheat. Nat-Biotechnology,1996,14:875-879
129.Barro F, Rooke L, Bekes F et al. Transformation of wheat with high molecular
weight subunit genes results in improved function properties. Nat Biotechnol 1997
Nov,15(12):1295-1299
130.Shewry P.R., and Hatham N,G.. Cereal seed storage protein:structures, properties
and role in grain utilization. J. Exp. Bot. 2002, 53(370):947-958
131.J.Sambrook, E.F.Fritsch, T.Maniatis. 金冬雁, 黎孟枫译. 分子克隆（第二版）,北
京: 科学出版社, 1995
132.Xu Shu-Ping( 徐 淑 平 ),WEI Zhi-Ming( 卫 志 明 ).Introduction to Method of
Microprojectile Bombardment and Its Application.plant physiology
参考文献 65
communication(植物生理学通讯)，1998，34：41-43
133.Papp AC, Snyder PJ, Sedra M, et al.Strategies for amplification of trinucleotide
repeats:optimization of fragile X and androgen receptor PCR. Mol Diagn,
1996,1(1)∶59
2001
2. 刘良式等. 植物分子遗传学. 1997，492
3. 刘良式等. 植物分子遗传学. 1997，210
4. 刘昱辉，王志幸，贾仕荣. 拟南芥 profiling 启动子 5’端缺失对维管束特异表达
的影响. 科学通报, 2001，5:835-828
5. 吴标，潘瑞琴，芦睿，田颖川. 竹节花黄斑驳病毒启动子的缺失分析及功能. 微
生物学报, 1999，39 (1):15- 21
6. 蒋浩，秦红敏，虞红梅. 笋瓜韧皮部蛋白基因启动子的克隆及功能初探. 农业生
物技术学报，1999，7 (1): 63-68
7. 黄伟达，姜学军，黄皓，贺斌等. 小麦高分子量谷蛋白基因转录调控相关的顺反
式相互作用的初步研究. 植物学报，1995，37:425-431
8. 李胜国，刘玉乐，康良仪，田波. 烟草花药特异性启动子的克隆、活性测定及雄
性不育基因和恢复基因的构建. 农业生物技术学报，1993，3 (3): 25-32
9. 杨洪全，卫志明，许智宏. Rch10 启动子引导 iaaL 基因在转荃因烟草中的表达导
致花器官发育的变异. 植物学报，1996，38 (7):541 – 547
10. 范惠琴，姚泉洪，黄晓敏，彭日荷. 矮牵牛查尔酮黄烷酮异构酶（chiA）基因启
动子的克隆和测序. 上海农业学报, 1998，14(3):14-16
11. 叶梁，李极，宋艳茹. 双价、三价种子特异性表达载体的构建及表达 PHB 合成
相关基因的转基因油菜的获得. 科学通报，2000，45(5):516
12. 赵友梅，王淑俭. 高分子量麦古蛋白亚基的 SDS-PAGE 图谱在小麦品质研究中
的应用. 作物学报，1990，16(3):208-218
13. 王瑞等. 一些优质小麦及其杂种后代高分子量谷蛋白亚基组成与面包品质之关
系. 西北农业学报, 1995，4:25－30
14. 刘香利. 小麦醇溶蛋白和高分子量麦谷蛋白亚基组成与品质关系研究，[硕士论
文] ，1999
15. 徐虹. 中间偃麦草麦谷蛋白亚基的鉴定、基因克隆与分子特征分析, [博士论文]，
2003
16. 王关林，方宏筠主编. 植物基因工程. 科学出版社，北京 2002，143-172
17. 解瑞立，万永芳，张慧，王道文. 多倍体山羊草物种中高分子量麦谷蛋白亚基组
成与其编码基因的分子克隆. 科学通报，2000，45(17):1867-1874
18. 张晓东，李冬梅，徐文英等. 利用基因枪将 HMW 谷蛋白亚基基因与除草剂 Basta
抗性基因导入小麦不同外植体获得转基因植株。遗传 HEREDITAS（Beijin）（增
56 小麦胚乳特异性启动子的克隆及高分子量谷蛋白 14 亚基基因的原核表达
刊），1998，20:3－8
19. 陈梁鸿，王新望，张晓东等. 小麦编码高分子量谷蛋白亚基基因的转化. 作物学
报，1999，25(4):437－440
20. 唐凤兰等. 利用花粉管通道法向小麦中导入麦谷蛋白高分子量优质亚基基因的
研究初报. 黑龙江农业科学，2002，3:1－2
21. 赵虹. 小麦基因枪转化体系的建立和抗除草剂基因的导入, [硕士论文]， 2001
22. 邓志勇. 高分子量麦谷蛋白 14 和 15 亚基的纯化及部分生化特性的研究, [硕士论
文] ，1999
23. 范三红. 小麦高分子量麦谷蛋白 14 和 15 亚基的研究, [硕士论文]， 2001
24. Donal, R,G. kand cashmore, A.R. RG Donald and AR Cashmore. Mutation of
either G box or I box sequences profoundly affects expression from the Arabidopsis
rbcS-lA promoter. EMBO J (1990) 9: 1717-1726
25. P Puente, N Wei, and XW Deng. Combinatorial interplay of promoter elements
constitutes the minimal determinants for light and developmental control of gene
expression in Arabidopsis. EMBO J (I996) 15: 3732-3743
26. Hawk-Bin Kwon, Soo-Chul Park. Identification Of a Light Responsive Region of
the Nuclear Gene Encoding the B Subunit of Chloroplast Glyceraldehyde
3-Phosphate Dehdrogenase from Arahidopsis thaliana. Plant Physiol(1994)
105:357-367
27. Slogan JS, Schwartz BW, Becker WM: Promoter analysis of a light of a
light-regulated gene encoding Hydroxyyruvate reductase, an enzyme of the
photorespi -ratory glycolate pathway. Plant J(1993)3:867-874
28. Steven G. Daniel, Wayne M. Becker. Trangenic analysis the 5'-and 3'-flanking
regions of the NADH-dependent hydroxypyuvate redutctase gene from Cucumis
sativus L. Plant-molecuar-biology (1995) 28:821-836.
29. Lagrange,T.,B. Axeloss,M. Structure and expression of the nuclear gene coding for
the chloroplast ribosmal protein L2: Developmental regulation of a house-keeping
gene by altemative promoters. MoI. Cell. Biol (1993) 13:2614-2622
30. Harrak, H. Largrange, T. The expression of nuclear genes encoding plastid ribosomal
precedes the experssion of chloroplast genes during early phases of chloroplast
development. Plant physiol(1995)108:685-692
31. Zhou,D.-X. Li,Y-F. Sequence specific interaction between SIF, a spinach nuclear
factor,and a negative cis-element conserved in plastid-related genes. J. Biol. Chem
(1992) 267:23515-23519
32. Thiery Lagrange, Syiviane Gauvin. S2F, a leaf-Specic trans-Acting Factor, Binds to a
参考文献 57
Novel cis-Acting Element and Differentially Activates the RPL21 Gene. The plant
Cell(1997) 9:1469-1479
33. Ke11er,B. Baumgartner, C. Vascualr-specific expression of the bean GRP1.8 gene is
negatively regulated. Plant cell (1991) 10:1051-1061
34. Hatton,D.Sablowski,R.,Yung M H ,et al. Two classes of cis sequences contribute to
tissue -specific expression of a PAL2 promoterr in transgenic tobacco.The plant
J(1995)7(6):859-876
35. Yin Y，Chen L，Beachy R.1997.Promoter elements required for phloem-specific gene
expression from the RTBV promoter in rice.Plant J.12(5):1179-88
36. Graham MW，Craig S，Waterhouse PM.1997.Expression patterns of vascular -specific
promoters Ro1C and Sh in transgenic potatoes and their use in engineering
PLRV-resistant plants.Plant Mol Biol.33(4):729-35
37. Koltunow,AN.,et al .Different Temporal and Spatial Gene Expression Patterns Occur
during Anther Development .Plant Cell(1990)2:1201-1224.
38. Schrauwen JAM，de Groot PFM，van Herpen MMA，van der Lee T，Reynen WH，
Weterings KAP，Wullems GJ.1990.Stage-related expression of mRNAs during pollen
development in lily and tobacco.Planta，182:298-304
39. Hanson DD ， Hamilton DA ， Travis JL ， Bashe DM ， Nascarenhas JP.
1989.Characterization of a poolen-specific cDNA clone from Zea mays and its
expression. Plant Cell,1:173-179.
40. Zou J-T, Zhan X-Y, Wu H-M, Wang H, Chueng AY. 1994.Characterization of a rice
pollen-specific gene and its expression.American Journal of Botany，81:552 –561
41. Pe ME, Frova C，Colombo L，Bineli G，Mastroianni N，Tarchini R，Visioli
G.1994.Molecular cloning of genes expressed in pollen of Sorghum bicolor.
Maydica，39:107-113
42. Batanero E ， Villaba M ， Lopez-Otin C ， Rodriguez R.1994.Isolation and
characterization of an olive allergen-like protein from lilac pollen.Sequence analysis
of three cDNA encoding protein isoforms.Eur J Biochem，221:187-193
43. Lombardero M.，Barbas JA，Moscoso del Prado J，Carreira J，1994.cDNA sequence
analysis of the main olive allergen.Olee I.Clin Exp Allergy，24:765-770
44. Twell D，Yamaguchi J，McCormick S，1990.Pollen-specific gene expression in
transgenic plants:Coordinate regulation of two different tomato gene promoters
during microsporrgenesis.Development，109:705-713
45. Green PJ.Yong MH Cuozzzo M ， kano-Murakami Y ， Silverstein P. ， Chua
N-H.1988.Binding site requirements for pea nuclear protein factor GT- correlate with
58 小麦胚乳特异性启动子的克隆及高分子量谷蛋白 14 亚基基因的原核表达
sequences required for ligft-dependent transcriptional activation of the rbcS-3A gene.
EMBO J, 7:4035-4044
46. Kuhlemeier C ， Cuozzo !M ， Green PJ ， Goyvaerts E ， Ward K ， Chua
N-H .1988.Localition and conditional redundancy of regulatory elements in rbcS-3A，
apea gene encoding thr small subunit of ribulose-bisphospgate carboxylase.Proc Natl
Acad Sci USA，85:4662-4666
47. Twell D，Yamaguchi J，Wing R，McCormick S，1991.Promotei analysis of three
genes that are coordinately expresse during pollen development reveals pollen
specific enhances sequences and shared regulatory elements.Genes Devel.，5:496-507
48. Eadl Y，Curie C，McCormick S.Pollen specificity elements reside in 30bp of the
proximal promoters of two polloen expressed genes.Plant Cell，7:373-384
49. N.Bate&D.TWell.1998.Functional architecture of a late pollen
promoter:pollen-specific transcription is developmentaaly regulated by multiple:tage
specific and dependent activator elements .Plant Molecular Biology，37:859-869
50. De Pater,S.,Pham,K. J. A 22-by Fragment of the Pea Lectin Promoter Containing
Essential TGAC-like Motifs Confers Seed-Specific Gene Expression .Plant cell
(1993)5:877-886.
51. Bustos,M.M.,Vegum,D. Positive and negative cis-acting DNA domains are required
for spatial and temporal regulation of gene expression by a seed storage protein
promoter .EMBOJ(1991)10:1469-1479
52. Marzable,P,Busk,P,K Ludevid,M.D .and Torrent,M. The bifactorial endosperm box
of r-zein gene:characterization and functional of thePb3 and GZM scis-acting
elements. Plant J(1998).16:41-52
53. Vicen-Carbajosa,J.,Onate,L.,Lara,P.,Diaz,L.and Carbonero,P.,Barly BLZ1:A bZIP
transcriptional activator that interact with endosperm-specific gene promoters. Plant
J(l998)13:629-640
54. Takaiwa,F.,Yamanouchi,U.,Yoshihara,T.,Washida,H.,Tanabe,F.,Kato,A.
Characterization of common cis-regulatory elements responsible for the
endosperm-specific expression of members of the rice glutelin multigene family.
Plant MOL.Biol(1996)3:1207-1221
55. Washida,H.,Wu,C.-Y,Yamanouchi,U.,akihanma,T.,Harada,K.and Takaiwa,F.
Identification of cis-regulatory elements required for endosperm expression of the
storage protein glutelin gene Glub-1. Plant MOL.Biol(1999)40,1-12
56. Zheng,Z,Kawagoe,Y,xiao,S,Li,Okita,T,Hau,TL,Lin,A.and Murai,.N. 5’distal and
proximal cis-acting regulatory elements are riquired for developmental control arice
参考文献 59
seed storagegene. Plant J(1993)4:357-366.
57. Yoshihra,Twashida,H.and Takaiwa,F.A 45-by proximal regioncontaining AACA and
GCN4 motif is suffient to confer endosperm-specific expression of the rice storage
protein glutein gene Glu-3. FEBS Lett(1996)383:213-218
58. Albani,D.,Hammond-Kosack,M.C.U., The wheat transcriptional activator SPA:a
seed-specific bZJP protein that recognizes the GCN4-like motif in the bifactoral
endosperm box of prolamin genes .Plant Cell(1997) 9:171-184
59. Mena,M.,Vicente-Carbajosa,J.,Schmidt,R.J. and Caarbonero,P. An
endosperm-specific DOF protein from barly,highly conserved in wheat ,binds to and
activies transciption from the prolamin -box of a native B- hordein promoter in barly
edospeerm box. Plant J(1998)16:53-62
60. Suzki,A.,Wu,C.-Y Washida,H.and Takaiwa,F. Rice NM protein OSMYB5
specifically binds to the AACA motif conserved among promoters of gene for storage
protein gllutelin. Plant cell Physiol(1998)39:555-559.
61. Schmidt,R.,Katagiri,M. Opaque-2 is a transcriptional activator that recognizes a
specific targert site in 22kD zein genes.Plant cell(1992) 4:689-700.
62. Wu,C.-YSuzuki,A.,Washida,H.and Takaiwa,F.The GCN4 motif in a rice glutilin gene
is essential for endosperm-specific gene expression and is actived by Opaque-2 in
transgenic rice plants. Plant J(1998)14,673-683
63. Chuan-Yin Wu,Haruhiko Washida.Quantitatve nature of the Prolamin-box,ACGT
and AACA motifs in a rice glutelin gene promoter:Minimal cis –element riquiremints
for endosperm-specific gene.The plant Journal (2000)23:415-421
64. Washida,H.,Wu,C.-Y,Yamanouchi,U.,akihanma,T.,Harada,K.and Takaiwa,F.
Identification of cis-regulatory elements required for endosperm expression of the
storage protein glutelin gene Glub-1. Plant MOL.Biol(1999)40,1-12
65. Robert L S,Thompson R D,Flavell R B.Tissue-specific expression of a wheat high
molecular weight glutenin gene in transgenic tobacco.Plant Cell,1989,1:569-578
66. Bate，N.，Spurr，C.，Forter，G.D.and Twell，D.1996. Maturation 一 specific
translational enhancement mediated by the 5，-UTR of a late pollen transcript. Plant
J.10:613-623
67. D. Hatton，R.Sablowski，M-H Yung，G.Smith，W. Schuch and M. Bevan.1995.Two
classes of cis sequences contribute to tissue-specific expression of a PAL2 promoter
in transgenic tobacco.The Ptant J.，7(6):859-876
68. Bronstein I, Martin CS, Fortin JJ, et al. Chemiluminescence: sensitive detection
technology for reporter gene assays［J］. Clin Chem,1996,42(9): 1542.
60 小麦胚乳特异性启动子的克隆及高分子量谷蛋白 14 亚基基因的原核表达
69. Koziel M. G.，BelandG.L.，Bowman C.，N.B.Carozzi，R.Crenshaw, et al，1993.Field
performance of elitee transgenic maize plants expressing an insecticidal protein
derived from Bacillus thuringiensis. Bio/Technology，11:194-200
70. Oxtoby，E.，Hughes，M.A.，1990.Engineering herbicide tolerance into crops.Trends
Biotechnol.8:61-65
71. Edwards，J.W.，Waiker，E.L.，Coruzzi，G.M.1990.Cell-specific expression in
transgenic plants reveals non-overlapping roles for chloroplast and cytosolic
glutamine synthetase .Proc.Natl.Acad.Sci.USA，87:3459-3463
72. Mol，J.，Stuitje，A.，van der Krpl，A.，Jorgensen，R.1989. Saying it with
genes:Molecular flower breeding.Trebds Biotechnol.7:148-153
73. Tunen A, Mur L, Mol JNM. 1990. Pollen- and anther-specific chia promoters from
petunia:Tandem promoters regulation of the chia gene. The Plant Cell, 2:393-401
74. Sato ， T. ， Theologis ， A. 1989.Cloning the mRNA encoding
1-ammminocyclopropane-1-carboxylate synthase, the key enzyme for ethylene
biosynthesis in plants.Proc.Natl.Acad.Sci .USA. 86:6621-662
75. Edwards，J.W.，Waiker，E.L.，Coruzzi，G.M.1990.Cell-specific expression in
transgenic plants reveals non-overlapping roles for chloroplast and cytosolic
glutamine synthetase .Proc.Natl.Acad.Sci.USA，87:3459-3463
76. Vandekerckhove，i.，Damme，i.F.，Lijsebettens，M.，Botterman，i.，De-Block，
M.，1989.Enkephalins produced in trangenic plants using modified 2S seed storage
proteins.Biotechnoloav.7:929-932
77. Firek S.，Draper J，Owen MRL.Gandecha A.，Cockburn W.，Whitelam GC. Secretion
of a functional single 一 chian Fv protein in transgenic tobacco plants and cell
suspension.1993
78. Ma JK-C，Hiatt A.，Hein M.，Vine ND.，Wang F.，Stab ila P.，van Dolleweerd C.，
Mostov K.，Lehner T.，1995.Generation and assembly of secretory antibodies in
plants.Science，268:716-719
79. Mason HS，Lam DM-K，Arntzen CJ. 1992.Expression of hepatitis B surface antigen
in transgenic plants.Proc Natl.Acad.Sci.USA, 89:11745-11749
80. Haq TA，Mason H. S.，Clements JD.，Arntzen CJ.，1995.Oral immunization with a
recombinant bacterial antigen produced in transgenic plants. Science, 268:714-716
81. Matsumoto S，I kura K，Ueda M，Sasaki R.1995.Characterization of a human
glycoprotein(erythropoietin)produced in cultutred tobacco cell，.Plant Mol.Biol.
27:1163-1172.
82. Ottoboni LMM，Lei te A，Yunes JA，Targon MLPN，de Souza Filho GA, Arruda P.，
参考文献 61
1993. Sequence analysis of 22kDa-like a-coixin and their comparison with
homologous zein and kafirin genes reveals highly conserved protein structure and
regulatory elements.Plant Mol Biol.，21:765-778
83. Adilson L，Edson L.K，Marc i o J.da S，Augusto D.L.，Rodrigo M. P .S.，Eric D .B.，
Hamza F.E-D.，Paulo A.，2000.Expression of correctly processed human growth
hormone in seeds of transgenic tobacco plants.Molecular Breeding，6:47-53
84. Wahlund KG, Gustavsson M, MacRitchie F, Nylander T,Wannerberger L(1996) Size
characterization of wheat proteins, particularly the gluteins, byasymmetrical flow
field-flow fractionation. J. Cereal Science 23:113-119
85. Stevenson SG, Prestom KR(1996) Flow Field-Flow Fractionation of Wheat Protein
Cereal Sci. 23:121-131
86. Wrihley CW(1996) Giant proteins with flour power. Nature 381:738-739
87. Seilmeier W, Belitz HD, Wieser H. Separation and quantitative determination of
high-molecular-weight subunit of glutenin from different wheat varieties and genetic
variants of the variety Sicco. Zeitschrift fur Lebensmittel Untersuchung
und-Forschung ，1991，192: 124-129
88. Weegles Pl, Hamer RJ, Schofield JD(1995) RP-HPLC and Capillary Electrophoresis
of Subunits from Glutein Isolated by SDS and Osborne Fraction. J Cereal Sci.
22:211-224
89. Vasil V,Castillo A M,Fromm M E and Vasil I K. Herbicide resistant fertile
transgenetic wheat plants obtained by mircoprojectile bombardment of regenerable
embyogenic callus, Bio/technology. 1992,10:667-674
90. Fredy Altpter,Vimla vasil,Vibbas Srivastava and Indra K Vasil. Intergration and
expression of the High-Moleclure-weight lutenin subunits 1AxI gene into wheat
Nature Biotechnology 1996, 14:1155-1159
91. Ann E Bkchl and Olin d Anderson . Expression of a Novel High-Molecular -weight
Glutenin subunit gene in transgenic wheat. Nadture Biotechnology 1996,14:875-879
92. Rooke L,Barro F,Tatham A S,Fido R,Steel S,Bekes F,Cras P,Martin A,Lazzeri P
A,Shewry P R, Barccelo P. 1999,Altered functional properties of tritordeum by
transformation with HMW glutein subunit genes.Theoretical And Applied Genetics.
99:851-859
93. Payne PI, Holt LM, Lawrence GJ(1983) Detection of a novel high molecular weight
subunit of glutenin in some Japnese hexaploid wheats. J. Cereal Sci. 1:3-8
94. Payne PI, Corfield KG, Blackman JA(1981) correlation between the inheritance of
certain high-molecular-weight subunit of glutenin and bread-making quality in
62 小麦胚乳特异性启动子的克隆及高分子量谷蛋白 14 亚基基因的原核表达
progenies of six corsses of bread wheat. J. Sci. Food Agric. 32:51-60
95. Anderson,O.D.and F,C.Greene,The characterization and comparative analysis of high
molecular weight glutenin genes from genomes A and B of a hexaploid bread
wheat.Theor.Appl.Genet. 1989b ,77:689－700
96. Anderson,O.D.F.C.Greene.Nudeotide sequences of the two high－molecular weight
glutenin genes from the D － genome of hexaploid wheat,Triticum aestivum
L.CV.cheyene.Nucleic Acids Res. 1989a ,17:461－462
97. Anderson,O.D.Litts.J.C. Nucleic acid sequence and chromosome assignment of a
wheat storage protein gene.Nucleic Acids Res 1984,12:8129－8144
98. Forde,J.et al, The nucleotide sequence of a HMW glutenin subunit gene located on
chromosome 1A of wheat (Triticum aestivum L.). Nucl.Acids Res. 1985,13:6817－
6832
99. Halford ,NG The mucleotide and deduced amino acid sequences of an HMW glutenin
subunit gene from chromosome 1B of bread wheat (Triticum aestivum L.)and
comparison with those of genes from chromosomes 1A and 1D.Theor Appl Genet
1987,75:117－126
100. Shenry,P.R.et al 1992,High molecular weight subunits of wheat glutenin J.Cereal
Sci 15:105－120
101.Deng ZY, Zhao HX, Fan SH, Ji WQ, Guo AG, Xue XZ. Purification and
biochemical characterization of high-molecular-weight-glutenin subunits 14 and
15.Yi Chuan Xue Bao.; 2001,28(1):46-51. Chinese
102.Mackie AM, Sharp PJ, Lagudah ES. The Nucleotide and Derived Amino Acid
Sequence of a HMW Clutenin Gene from Triticum tauschii and Comparison with
those from the D Genome of Bread Wheat. Journal of Cereal Science,1996, 24:73-78
103.Bustos ad, Rubio P, Jouve N. Molecular characterization of the inactive allete of the
gene Glu-AI and the development of a set of AS-PCR markers for HMW glutenins of
wheat. Theor. Appl. Genet.2000,100: 1085-1094
104.Wan Y, Wang D, Shewry PR, Halfordb NG(2002) Isolation and characterization of
five novel high molecular weight subunit of glutein genes from Triticum timopheevi
and Aegilops cylindrical. Theor Appl Genet 104:828-839
105.Shewry P.R., Napier J.A., Tatham A.S.. Seed storage proteins: structures and
biosynethsis.The plant cell., 1995,7:945-956
106.Shewry P.R., and Halford N.G.. Cereal seed storage protein:structures, properties and
role in grain utilization. J. Exp. Bot. 2002, 53(370):947-958
107.Roberts L S., Thompson R.D., Flavell R.B. Tissue-specific expression of a wheat
参考文献 63
high-molecular-weight glutenin gene in transgenic tobacco. Plant cell.1989.
1:569-578
108.Ballesteros J, Alvarez B, Gimenez J, Ramirez C, Cabrera A, Martin A. Introgression
of 1Dx5+1Dy10 into Tritordeum. Theor Appl Genet. 2003. 106(4):644-8
109.Rooke L, Barro F, Tatham AS, Fido R, Steele S, Bekes F, Gras P, Martin A, Lazzeri
PA, Shewry PR, Barcelo P. Altered functional properties of tritordeum by
transgprmation with HMW glutein subunit genes. Theor Appl Genet. 1999a.
99:851-858
110.Sangtong V., Moran R., Chikwamba R., Wand K., Woodman-Clikeman, Long M.J.,
Lee M., Scott M.P. Expression and inheritance of the wheat Glu-1Dx5 gene in
transgenic maize. Theor Appk Genet. 2002. 105:937-945
111.Bartel D,Thompson R D, Rodthstein S. 1985 Synthesis of a wheat stroge protein
subunit in Eschcerichia coliusing Novel expression vector,Gene,35(1-2):157-167
112.Maruyama N,Ichisc K,Kasube T,Kishimoto T,Kawase S,Matsumura Y,Takeuchi
Y,Sawada T,Utsumi S.1998,Identification of major wheat allergens by of the
Escherichia coli expression system Eur J Biochem Aug 1,255 (3):739-745
113.Calili J M,Rousset M,Qualate C Q.Kasarda D D .1990.Use of recmbinant inbred
lines of wheat for study of associationsof high-molecular-weight glutenia subunit
alleles to quartitative traits I.Grain yield and quality predication tests. Theoretical
And Applied Genetics.79:321-330
114.Anderson OD, Greec FC,Halford NG,Shewry PR,Malpica-Romero J-M 1989
Nucleotide sequence of die two high-molecular-weight glutenin gene from thr
D-genome of a bexaploid bread wheat, Triticum aestivum L.ev. Cheyenne. Nucleic
Acid Research 17:461-462.
115.Coraggio I,Compagno C,Martegani E,Ranzi B M,Sale E,Alberghina L and Viotti A.
1986 Transcription and expression of zein sequence in yeast under natural plant or
yeast promters. EMBO Journal,5:459-465
116.Campagno C,Coraggio J,Ranzi B M,Alberghina A L,Viotti A and MarteganiE.
1987,Translation of regulation of expression of zein cloned in yeast under inducible
GAL promoter.Biochemical and biophysical research commeication,146:809-814
117.Neill J D,Litts J C,Andasan O D,Greene F C and Stiles J I.1987,Expression of a
wheatα-gliadin gene in Soccharomcyces carevisiae, Gene,55:303-317
118.Scheets K and Hedgeoth C. 1989.Expression of a wheat α-gliadin gene in
Saccharomcyces cerevisiae from a yeast ADH1 promoter. Journal of agricultural and
food chemistry.37:829-833
64 小麦胚乳特异性启动子的克隆及高分子量谷蛋白 14 亚基基因的原核表达
119.Bartel D,Altosaar I,Harbert N P,Barker R F and Thompaon R D.1986, molecular
analysis of γ-gladin gene families at yhe complex Gli-I locus of bread wheat
(T.aestivum L.).uTheoretical and applied Genetics.72:845-853
120.Miyamoto C,Smithe G E,Farred-Towt J,Chizzonile R,Summers M D and Ju G.
1985,Productin of human c-myc protein in insect cells infected with a baculovirus
expression vector. Molecular and cell biology .5:2860-2865
121.Emery V C aad Bisbop D H L,1987.The development of multiple expression vector
for high level synthesis of eukaryotic proteins:expression of LCMVN polyhedrin
protein in a recombinant baculovirus system. Protein Engineer,1:359-366
122.Matsuura Y,Posse R D,Overton H A,and Bioshop D H L.1987,Baculovirus
expression vectors:The requirements for high level expression of proteins ,including
glycoproteins, Journal of General Virology ,68:1233 –1250
123.Ng P K W and Busluk W.1989,Concerning the nomenclature of high moleculare
weight glutein subuntits. Journal of cereal science,9:53-60
124.Lee Y.K., Bekes F., Ciaffi M. The low molecular weight glutenin subunit protenins
of primitive wheats. Ⅳ. Functional properties of products from individual genes.
Theor Appl Genet 1999c. 98:149-155
125.Sissons M.J., Bekes F., Skerritt J.H. 1998. Isolation and Functionality testing of low
molecular glutein subunits Cereal Chem., 75(1):30-36
126.D’ovidio R., Anderson O.D., Madci S., Skeritt J. and Porecddu E. Construction of
novel wheat high-Mr glutein subunit gene variability:modification of the repetitive
domain and exression in E.coli. Journal of cereal science. 1997b.25:1-8
127.Anderson O.D., Kuhl J C., Tam A. Construction and expression of a synthetic wheat
storage protein gene.Gene. 1996. 174:51-58
128.Blechl A E,Anderson O D. Expression of a novel high-molecular-weight glutenin
subunit gene in transgenic wheat. Nat-Biotechnology,1996,14:875-879
129.Barro F, Rooke L, Bekes F et al. Transformation of wheat with high molecular
weight subunit genes results in improved function properties. Nat Biotechnol 1997
Nov,15(12):1295-1299
130.Shewry P.R., and Hatham N,G.. Cereal seed storage protein:structures, properties
and role in grain utilization. J. Exp. Bot. 2002, 53(370):947-958
131.J.Sambrook, E.F.Fritsch, T.Maniatis. 金冬雁, 黎孟枫译. 分子克隆（第二版）,北
京: 科学出版社, 1995
132.Xu Shu-Ping( 徐 淑 平 ),WEI Zhi-Ming( 卫 志 明 ).Introduction to Method of
Microprojectile Bombardment and Its Application.plant physiology
参考文献 65
communication(植物生理学通讯)，1998，34：41-43
133.Papp AC, Snyder PJ, Sedra M, et al.Strategies for amplification of trinucleotide
repeats:optimization of fragile X and androgen receptor PCR. Mol Diagn,
1996,1(1)∶59