野生大豆基因在拟南芥中高通量表达技术研究
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摘要
低温、干旱和盐碱是制约我国乃至世界农业生产的重大问题,提高作物的耐低温、干旱和盐碱能力,对于提高粮食产量、开发耕地资源、解决我国粮食问题具有重大的战略意义。
近年来,随着分子生物学的飞速发展和转基因技术的日趋成熟,通过基因工程技术改良作物的抗渗透胁迫能力出现了良好的势头。阐明植物抗渗透胁迫的分子机理,从大量的渗透胁迫相关基因中找到“关键基因”,是作物抗渗透胁迫基因工程的重要前提。本研究通过构建与SMART技术相匹配的通用高效超量表达植物载体卡盒,研究并建立耐盐野生大豆全长cDNA超量表达文库的构建技术和抗渗透胁迫转基因植株筛选技术,为实现野生大豆基因在拟南芥中的随机超量表达,从中筛选抗渗透胁迫转基因植株并分离抗渗透胁迫基因奠定基础,为高通量、简便有效的筛选抗渗透胁迫基因探索一条新的途径。
本研究的主要研究成果如下:
1.与SMART技术相匹配的通用高效超量表达植物载体卡盒的构建
构建了表达框两侧翼带有核基质结合区(MAR),由E12启动子调控的含有SfiA、SfiB克隆位点的通用高效植物表达载体卡盒pEMT-5。pEMT-5适用于构建应用SMART技术获得的全长cDNA的超量植物表达载体或超量表达文库。
2.植物载体卡盒pEMT-5的功能验证
将载体pEMT-5与其不含MAR的原始载体pBHT-5同时转化烟草,通过半定量RT-PCR及实时荧光定量PCR在转录水平分析005基因的表达。并通过实时荧光定量PCR实验,进行数据分析,结果表明转化质粒pEMT-5的烟草植株内005基因的转录表达水平是转化质粒pBHT-5的烟草植株内005基因表达的3.89倍,进一步验证了半定量RT-PCR实验结果。
3.野生大豆全长cDNA超量表达文库构建技术的建立
运用Clontech SMART~(TM)技术获得了两端带有SfiA、SfiB酶切位点的野生大豆渗透胁迫条件下的全长cDNA,将全长cDNA克隆于通用高效植物表达载体卡盒pEMT-5。电击法转化大肠杆菌DH5α,经质粒酶切鉴定,其重组率为96.29%。并文库滴度测定实验,计算出扩增前文库滴度为1.96×10~8 cfu/mL,扩增后文库滴度为2.0×10~(10) cfu/mL。同时利用三亲杂交法将cDNA文库转化根癌农杆菌LBA4404,构建了全长cDNA超量表达农杆菌文库,用于Floral dip法侵染拟南芥。
4.基因文库Floral dip法转化拟南芥及转基因植株的获得
(1)将构建好的pEMT-cDNA文库,Floral dip法转化拟南芥,通过拟南芥的遗传转化获得转基因的种子,并收获拟南芥种子用于下一步的筛选试验。
(2)通过拟南芥种子发芽阶段km筛选压力实验,最后确定了拟南芥种子发芽阶段km的筛选压力为10mg/L。
(3)从约10000粒拟南芥种子中筛选出有Km抗性的110株,计算转化率约为1.1%。
Low temperature,salinity and drought are a major issue in our country and world agricultural production.Improving the tolerance of these abiotic stress of crops,is of great strategic significance for increasing food production,developing farming resources and solving the food problem.
In recent years,with the rapid development of molecular biology and the mature of transgenic technology,anti-modified crops through genetic engineering against osmotic stress.There has been a good momentum.Explaining the molecular mechanism of osmotic stress in plant and finding "key genes" from a large number of stress-related genes,is an important prerequisite for osmotic stress genetic engineering of crops.Our research aims at these problems,constructed plant expression vector generic card boxes with the matrix attachment regions,MAR.Taking the wild soybean of northeast as the material,established high-throughput gene over expression system,and the system of screening the transgenic plants against osmotic stress.Therefore,our study is of great significance for tolerance basic theoretical research,foundation of new technologies system,agriculture utilizing,and other areas.
Main research results are as follows:
1.Plant Expression Vector Construction
We had cloned P-MN and E-MN sequence including MAR sequence by overlap extension PCR method.At the same time,we constructed one plants common effective expression vector card boxes pEMT-5 with MAR and SfiA、SfiB restriction sites,promoted by E12.The vector is suitable for constructing over-expression library which is obtained by SMART technology.
2.Plant Expression Vector functional analysis
The vectors pEMT-5 and pBHT-5 were transferred into tobacco by Agrobacterium Tumefaciens.Semi-quantitative RT-PCR and Real-time PCR analysis the same 005 gene expressed in pBHT-5 and pEMT-5 vectors in transgene plants.With the analysis of transcription levels,005 expression levels in pEMT-5 are more 3.89 times than in pBHT-5.
3.Wild Soybean conditions of osmotic stress of a full-length cDNA clone
We obtained the full length cDNA of wild soybean under the conditions of osmotic stress by Clontech SMART~(TM) technique.Then,a full-length cDNA cloned the vector pEMT-5.The result of the analysis is that the primitive titer of the library was 1.96×10~8cfu/mL and the end titer of the library was 2.0×10~(10)cfu/mL,the recombination efficiency of the cDNA library was 96.29%by PCR identification.The library fits gene over-expression.
4.Gene library transferred into Arabidopsis and transgene plant were selected
(1) Gene library transferred into Arabidopsis by Floral dip and harvested Seeds.
(2) In order to select transgene seeds,Km selection pressure is 10mg/L.
(3) We harvested 10000 seeds,there are 110 Strains by Km selection pressure, transformation ratio 1.1%.
近年来,随着分子生物学的飞速发展和转基因技术的日趋成熟,通过基因工程技术改良作物的抗渗透胁迫能力出现了良好的势头。阐明植物抗渗透胁迫的分子机理,从大量的渗透胁迫相关基因中找到“关键基因”,是作物抗渗透胁迫基因工程的重要前提。本研究通过构建与SMART技术相匹配的通用高效超量表达植物载体卡盒,研究并建立耐盐野生大豆全长cDNA超量表达文库的构建技术和抗渗透胁迫转基因植株筛选技术,为实现野生大豆基因在拟南芥中的随机超量表达,从中筛选抗渗透胁迫转基因植株并分离抗渗透胁迫基因奠定基础,为高通量、简便有效的筛选抗渗透胁迫基因探索一条新的途径。
本研究的主要研究成果如下:
1.与SMART技术相匹配的通用高效超量表达植物载体卡盒的构建
构建了表达框两侧翼带有核基质结合区(MAR),由E12启动子调控的含有SfiA、SfiB克隆位点的通用高效植物表达载体卡盒pEMT-5。pEMT-5适用于构建应用SMART技术获得的全长cDNA的超量植物表达载体或超量表达文库。
2.植物载体卡盒pEMT-5的功能验证
将载体pEMT-5与其不含MAR的原始载体pBHT-5同时转化烟草,通过半定量RT-PCR及实时荧光定量PCR在转录水平分析005基因的表达。并通过实时荧光定量PCR实验,进行数据分析,结果表明转化质粒pEMT-5的烟草植株内005基因的转录表达水平是转化质粒pBHT-5的烟草植株内005基因表达的3.89倍,进一步验证了半定量RT-PCR实验结果。
3.野生大豆全长cDNA超量表达文库构建技术的建立
运用Clontech SMART~(TM)技术获得了两端带有SfiA、SfiB酶切位点的野生大豆渗透胁迫条件下的全长cDNA,将全长cDNA克隆于通用高效植物表达载体卡盒pEMT-5。电击法转化大肠杆菌DH5α,经质粒酶切鉴定,其重组率为96.29%。并文库滴度测定实验,计算出扩增前文库滴度为1.96×10~8 cfu/mL,扩增后文库滴度为2.0×10~(10) cfu/mL。同时利用三亲杂交法将cDNA文库转化根癌农杆菌LBA4404,构建了全长cDNA超量表达农杆菌文库,用于Floral dip法侵染拟南芥。
4.基因文库Floral dip法转化拟南芥及转基因植株的获得
(1)将构建好的pEMT-cDNA文库,Floral dip法转化拟南芥,通过拟南芥的遗传转化获得转基因的种子,并收获拟南芥种子用于下一步的筛选试验。
(2)通过拟南芥种子发芽阶段km筛选压力实验,最后确定了拟南芥种子发芽阶段km的筛选压力为10mg/L。
(3)从约10000粒拟南芥种子中筛选出有Km抗性的110株,计算转化率约为1.1%。
Low temperature,salinity and drought are a major issue in our country and world agricultural production.Improving the tolerance of these abiotic stress of crops,is of great strategic significance for increasing food production,developing farming resources and solving the food problem.
In recent years,with the rapid development of molecular biology and the mature of transgenic technology,anti-modified crops through genetic engineering against osmotic stress.There has been a good momentum.Explaining the molecular mechanism of osmotic stress in plant and finding "key genes" from a large number of stress-related genes,is an important prerequisite for osmotic stress genetic engineering of crops.Our research aims at these problems,constructed plant expression vector generic card boxes with the matrix attachment regions,MAR.Taking the wild soybean of northeast as the material,established high-throughput gene over expression system,and the system of screening the transgenic plants against osmotic stress.Therefore,our study is of great significance for tolerance basic theoretical research,foundation of new technologies system,agriculture utilizing,and other areas.
Main research results are as follows:
1.Plant Expression Vector Construction
We had cloned P-MN and E-MN sequence including MAR sequence by overlap extension PCR method.At the same time,we constructed one plants common effective expression vector card boxes pEMT-5 with MAR and SfiA、SfiB restriction sites,promoted by E12.The vector is suitable for constructing over-expression library which is obtained by SMART technology.
2.Plant Expression Vector functional analysis
The vectors pEMT-5 and pBHT-5 were transferred into tobacco by Agrobacterium Tumefaciens.Semi-quantitative RT-PCR and Real-time PCR analysis the same 005 gene expressed in pBHT-5 and pEMT-5 vectors in transgene plants.With the analysis of transcription levels,005 expression levels in pEMT-5 are more 3.89 times than in pBHT-5.
3.Wild Soybean conditions of osmotic stress of a full-length cDNA clone
We obtained the full length cDNA of wild soybean under the conditions of osmotic stress by Clontech SMART~(TM) technique.Then,a full-length cDNA cloned the vector pEMT-5.The result of the analysis is that the primitive titer of the library was 1.96×10~8cfu/mL and the end titer of the library was 2.0×10~(10)cfu/mL,the recombination efficiency of the cDNA library was 96.29%by PCR identification.The library fits gene over-expression.
4.Gene library transferred into Arabidopsis and transgene plant were selected
(1) Gene library transferred into Arabidopsis by Floral dip and harvested Seeds.
(2) In order to select transgene seeds,Km selection pressure is 10mg/L.
(3) We harvested 10000 seeds,there are 110 Strains by Km selection pressure, transformation ratio 1.1%.
引文
董英山,庄炳昌,赵丽梅等.2000.中国野生大豆遗传多样性中心[J].作物学报,26(5):521-527.
董艳珍,李关荣.2004.RNA干扰的研究进展[J].生物技术通讯,15(1):60-62.
代翠红,李杰,朱延明,纪巍,柏锡.2005.不同DNA提取方法对4种重要作物DNA提取效率的比较[J].东北农业大学学报,36(3):329-332.
傅作中,程远国.张玉静等.2002.用长距PCR法构建恶性疟原虫cDNA表达文库[J].热带医学杂志,2(3):225-229.
付绍红,牛应泽,杨洪全等.2004.表面活性剂silwetL-77对floral-dip转化甘蓝型油菜效果的影响[J].分子植物育种,2(5):661-666.
高健,许晓风,乌慧玲,等.2004.特异种质烟草全长cDNA文库的构建及质量分析[J].安徽农业大学学报,31(1):22-25.
关荣霞,刘秀敏,常汝镇等.2006.辽宁新宾县原位保护区野生大豆(Glycine soja Sieb.& Zucc.)遗传多样性分析[J].高技术通讯,16(1):67-72.
海林,王克晶.2002.半野生大豆种质资源SSR位点遗传多样性分析[J].西北植物学报,22(4):751-757.
何宝坤,王玉洁,孙立平.2005.蛋白质组学研究技术及其在植物抗渗透胁迫研究中的应用[J].生物技术通报,2:18-22.
纪巍,李杰,朱延明,柏锡,代翠红,侯爱菊.2005.不同启动子调控的DREB1A 基因对黄瓜的遗传转化[J].东北农业大学学报,36(4):442-447.
纪宗玲,刘继中,陈苏民.2002.基因功能的研究方法[J].生物工程学报,18:116-120.
来永才,林红,方万成等.2005.黑龙江省野生大豆优异资源筛选、评价及利用研究[J].中国农学通报,21(6):379-382.
李杰,李晶,朱延明,张晶,代翠红,纪巍,才华.2003.DREB1A基因植物表达载体的构建[J].东北农业大学学报,34(2):199-204.
李俊华,张艳春,徐云远等.2004.拟南芥室内培养技术[J].植物学通报,21(2):201-204.
李向华,王克晶,李福山等.2005.野生大豆(Glycine soja)研究现状与建议[J].大豆科学,24(4):305-309.
李向华,田子罡,李福山.2003.新考察收集野生大豆与已保存野生大豆的遗传多样性比较[J].植物遗传资源学报,4(4):345-349.
李秋莉,杨华等.2002.植物甜菜碱合成酶的分子生物学和基因工程[J].生物工程进展,22(1):84-86.
李旭刚,路子显,陈蕾等.2001.豌豆核基质结合区的分离及其在转基因烟草中的功能分析[J].中国科学(C辑),31(3):230-237.
梁慧敏,夏阳,王太明.2003.植物抗寒冻、抗旱、耐盐基因工程研究进展[J].草业学报,12(3):1-7.
刘西燕.2008.OsMAPK4基因水稻突变体的培育及功能分析.东北农业大学,43.
刘文华,王义良,陈慧萍等.2002.玫瑰红绿海葵触手cDNA表达文库的构建和初步分析[J].生物工程学报,18(6):749-753.
刘凡,王国英,曹鸣庆.2003.农杆菌介导的植物原位转基因方法研究进展[J].分子植物育种,1(1):108-116.
刘守伟,刘士勇.2007.拟南芥室内培养技术研究[J].东北农业大学学报,38(2):279-281.
刘峰,施卫明.2006.拟南芥室内水培方法的改进[J].土壤,38(1):102-105.
刘悦萍,宫飞,赵晓萌.2005.水杨酸介导的信号转导途径与植物抗逆性[J].中国农学通报,21(7):227-229.
罗会颖,姚斌,袁铁铮等.2004.来源于Escherichia coli的高比活植酸酶基因的高效表达[J].生物工程学报,20(1):78-84.
吕宪禹,卢茜,刘桂琴,等.2002.导入野生大豆DNA小麦后代的农艺性状研究[J].南开大学学报:自然科学版,35(4):103-105.
马艺沔.2003.利用转基因拟南芥研究质外体CaM在植物生长发育中的作用[D].毛新国,孔秀英,赵光耀等.2005.利用改进的Cap-trapper法构建拟斯卑尔脱山羊草(Ae.speltoides Tausch)全长cDNA文库[J].遗传学报,32(8):811-817.
萨姆布鲁克,拉塞尔.2002.分子克隆实验指南[M].黄培堂等译.科学出版社.
苏金,朱汝财.2001.渗透胁迫调节的转基因表达对植物抗旱耐盐性的影响[J].植物学通报,18(2):129-136.
陶爱林,林兴华,张端品.2003.获取全长cDNA若干方法的比较[J].武汉植物学研究,21(2):179-186.
田佳.2008.拟南芥渗透胁迫应答基因的筛选及功能分析.东北农业大学,42.
涂艳阳,徐如祥,杨志林等.2003.人脑瘤组织全长cDNA文库的构建[J].第一军医大学学报,23(9):916-920.
王秀玲,卢茜,刘君等.2003.野生大豆DNA导入小麦及RAPD分子验证[J].南开大学学报:自然科学版,36(2):37-40.
王皓,康现江,王琦.2007.重组PCR技术研究进展和应用[J].中国生物工程杂志,27(5):153-156.
王曙明,孙寰,王跃强等.2002.大豆杂种优势及其高优势组合选配的研究:F1代籽粒产量的杂种优势与高优势组合选配[J].大豆科学,21(3):161-167.
王敏,刘萍,石明旺等.2005.野生大豆种子cDNA文库的构建与分析[J].生物技术通讯,16(5):509-511.
王爱荣,刘丽,周洁等.2005.拟南芥室內简易栽培和田间大规模种植的方法[J].福建农林大学学报:自然科学版,34(2):252-254.
王克晶,李福山.2000.我国野生大豆(Glsoja)种质资源及其种质创新利用[J].中 国农业科技导报,2(6).69-72.
王关林,方宏筠.2002.植物基因工程原理与技术[M].北京:科学出版社,473-474.
王天云,柴玉荣,袁保梅等.2004.核基质结合区对转基因表达的影响及其作用机制[J].细胞生物学杂志,26(6):587-590.
王艺磊,张子平.2003.日本对虾精巢和卵巢全长cDNA文库的构建[J].动物学杂志,38(2):9-13.
吴才君,范涉英.2004.植物转基因沉默[J].江西农业大学学报:155-158.
徐光硕,饶勇强,陈雁等.2004.用in planta方法转化甘蓝型油菜[J].作物学报,1:1-5.
徐春晖,夏光敏,贺晨霞.2002.农杆菌转化系统研究进展[J].生命科学,4(4):224-225.
许亮,卢向阳,田云.2003.后基因组时代基因功能分析的策略[J].中国生物工程杂志,23:29-34.
许志茹,李玉花.2004.基因芯片技术在植物研究中的应用[J].生物技术,14(6):70-72.
杨洪强,梁小娥.2003.高等植物中的CDPK及其生理生化功能[J].山东农业大学学报(自然科学版),34(2):284-288.
杨光宇,王洋,马晓萍等.2005.野生大豆种质资源评价与利用研究进展[J].吉林农业科学,30(2):61-63.
袁盛凌,段海清,张兆山.2003.蛇类凝血酶Calobin cDNA的设计合成与克隆[J].生物技术通讯,14(5):357-360.
于玲,王莱,牛吉山,陈佩度.2003.植物功能基因组研究进展[J].西北师范大学学报,39:104-116.
张可伟,王健美,郑成超.2004.核基质结合序列(MAR)与基因表达调控[J].生物工程学报,20:6-9.
张震祯,徐煜泉,黄海.2006.拟南芥-一把通往生命科学奥秘大门的钥匙[J].生命科学,18(5):442-446.
赵洪锟,王玉民,李启云等.2001.中国不同纬度野生大豆和栽培大豆SSR分析[J].大豆科学,20(3):172-176.
赵团结,盖钧镒.2006.大豆不育细胞质资源的发掘与鉴定[J].作物学报,32(11):1604-1610.
周晓馥,庄炳昌,王玉民等.2002.利用RAPD与SSR技术进行野生大豆种群内分化的研究[J].中国生态农业学报,10(4):629.
庄炳昌.1999.中国野生大豆研究二十年[J].吉林农业科学,24(5):3-10.
朱维岳,周桃英,钟明等.2006.基于遗传多样性和空间遗传结构的野生大豆居群采样策略[J].复旦大学学报:自然科学版,45(3):321-327.
Allen GC,Spiker S,Thompson WF.2000.Use of matrix attachment regions(MARs)to minimize transgene silencing[J].Plant Mol Biol,43(223):361-376.
Ahre n D, Tholander M, Fekete C, et al. 2005. Comparison of gene expression in trap cells and vegetative hyphae of the nematophag ous fungus Monacrosporium haptotylum [J]. Microbiology, 51 (3) : 789-803.
Brouwer C, Bruce W, Maddock S, et al. 2003. Suppression of transgene Silencing by Matrix Attachment Regions in Maize. A dual Role for the Maize 5'-ADHl Matrix Attachment Region[J]. Plant Cell, 14: 2251-2264.
Butaye KM, Goderis IJ, Wouters PF, et al. 2004. Stable high-level transgene expression in Arabidopsis thaliana using gene silencing mutants and matrix attachment regions [J]. Plant J, 39 (3) : 440-449.
Carninci P, Waki K, Shiraki T, et al. 2003 . Targeting a complex transcriptome: the construction of the mouse full-length cDNA encyclopedia [J]. Genome Research, 13: 1273-1289.
Chenchik A, Moqadam F, Siebert P. 1996. RNA: Isolation, Analysis and Synthesis [M]. New York: Wiley-Liss, 273-321.
Curtis IS, Nam HG. 2001. Transgenic radish (Raphanus sativus L. Iongipinnatus Bailey) by floral-dip method plant development and surfactant are important in optimizing transformation efficiency [J]. Transgenic Research, 10: 363-371.
Curtis IS. 2005. Production of transgenic crops by the floral-dip method[J]. Methods Mol Biol, 286: 103-106.
Clough SJ. 2005. Floral dip: agrobacterium-mediated germ line transformation [J]. Methods Mol Biol, 286: 91-102.
Cheng YX, Long M. 2007. A cytosolic NADP-malic enzyme gene from rice ( Oryza sativa L. ) confers salt tolerance in transgenic Arabidopsis [J]. Biotechnology Letters, 29 (7) : 1129-1134.
Chen M, Wang QY, Cheng XG, et al. 2007. GmDREB2, a soybean DRE-binding transcription factor , conferred drought and high-salt tolerance in transgenic plants [J]. Biochemical and Biophysical Research Communications, 353 (2) : 299-305.
Chen SH, Guo SL, Wang ZL, et al. 2007. Expressed sequence tags from the halophyte Limonium sinense [J]. DNA Sequence, 18 (1) : 61-67.
Dastidar K G, Maitra S, Goswami L, et al. 2006. An insight into the molecular basis of salt tolerance of L-my-oinositol 1 -Psynthase (PcINO 1) from Porteresia coarctata (Roxb. ) Tateoka, a halophytic wild rice [J]. Plant Phsiology, 140 (4) : 1279-1296.
Francois Chaumont, Menachem Moshelion, Mark J, Danielst. 2005. Regulation of plant aquaporin activity[J]. Biol Cell, 97: 749-764.
Francois T, Anne P, Pierre B, et al. 2005 . Enhancement of maize transformation efficiency by the use of maize matrix attachment regions [J]. Plant Cell, 80: 295-303.
Fu SF, Chou WC, Huang DD, Huang HJ. 2002. Transcriptional regulation of a rice mitogen-activated protein kinase gene , OsMAPK , in response to environmental stresses[J]. Plant Cell Physiol, 43 (8) : 958-63.
Fu XH, Huang YL, Deng SL, et al. 2005. Construction of a SSH library of Aegiceras corniculatum under salt stress and expression analysis of four transcripts [J]. Plant Science, 169 (1) : 147-154.
Glazko G V, Rogozin I B, Glazkov M V. 2001. Comparative Study and Prediction of DNA Fragments Associated with Various Element of the Nuclear Matrix [J]. Biochem BiophysActa, 1517: 351-364.
Gernot G. 2003. Mapping multiple consequenced T-DNA integration sites within the Arabidopsis genome[J]. Bioinformatics, 19 (5) : 579-586.
Guo XL, Cao YR, Cao ZY, et al. 2004. Molecular cloning and characterization of a stress-induced peroxiredoxin Q gene in halophyte Suaedasalsa [J]. Plant Science, 167(5) : 969-975.
Gao F, Gao Q, Duan X G, et al. 2006. Cloning of an H~+-PPase gene from Thellungiella halophila and its heterologous expression to improve tobacco salt tolerance [J]. Journal of Eeperimental Botany, 57 (12) : 3259-3270.
Guo SL, Yin HB, Zhang X, et al . 2006. Molecular cloning and characterization of a vacuolarn H~+-pyrophosphatase gene, SsVP, from the halophyte Suaeda salsa and its over-expression increases salt and drought tolerance of Arabidopsis [J]. Plant Molecular Biology, 60 (1) : 41-50.
Geng DG, Han Y, Wang YQ, et al. 2004. Construction of a system for the stable expression of foreign genes in Dunaliella salina [J]. Acta Bot Sin, 46 (3) : 342-346.
Girod PA, Zahn Zabal M, Mermod N. 2005. Use of the chicken lysozyme 5'matrix attachment region to generate high producer CHO cell lines [J]. Biotechnol Bioeng, 91 (1) : 121.
Gspar YM. 2004. Characterizati on of the Arabidopsis lysine-rich arabinogalactan protein AtAGP17 mutant (rat1) that results in a decreased efficiency of agrobacterium transformation [J]. Plant Physiol, 135 (4) : 2162-71.
Garcia Mata C. Lamattina L. 2001. Nitric oxide induces stomatal closure and enhances the adaptive plant response against drought stress[J]. Plant Physiol, 12 (6) : 1196-1204.
Hu HH, Dai MQ, Yao JL, et al. 2006. Over-expressing a NAM, ATAF, and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice [J]. Proc Nat Acad Sci U. S. A. , 103 (35) : 12987-12992.
Hmida-Sayari A, Gargouri-Bouzid R, Bidani A, et al. 2005. Over-expression of Delta ( 1 ) - pyrroline-5-carboxylate synthetase increases praline production and confers salt tolerance in transgenic potato plants [J]. Plant Science, 169 (4) : 746-752. Halweg C, Thompson WF, Spiker S. 2005. The rb7 matrix attachment region increases the likelihood and magnitude of transgene expression in tobacco cells: a flow cytometric study [J]. Plant Cell, 17 (2) : 418-429.
Ichikawa T, Nakazawa M, Kawashima M, el al. 2003. Sequence database of 1172 T-DNA insertion sites in Arabidopsis activation-tagging lines that showed phenotypes in T1 generation[J]. Plant J, 36 (3) : 421-429.
Jiang GZ, Lu YM, Niu XL, et al. 2005. The actin gene promoter driven bar as a dominant selectable marker for nuclear trans formation of Dunaliella salina [J]. Acta Genet Sin, 32 (4) : 424-433.
JiaJ, FuJ, Zheng J, et al. 2006. Annotation and expression profile analysis of 2073 full-length cDNAs from stress-induced maize (Zea mays L. ) seedlings [J]. Plant J, 48 (5) : 710-727.
KY amaguchi, Shinozaki K. 2003. OsDREB genes in rice, Oryza sativa L, encode transcription activators that function in drought, high-salt and cold responsive gene expression[J]. Plant J, (4) : 751-763.
Kurusu T, Yagala T, Miyao A, Hirochika H, Kuchitsu K. 2005. Identification of a putative voltage-gated Ca~(2+) channel as a key regulator of elicitor-induced hypersensitive cell death and mitogen-activated protein kinase activation in rice[J]. Plant, 42 (6) : 798-809.
KUWAYAMA H, OBARA S, MORIO T, et al. 2002. PCR-mediated generation of a gene disruption construct without the use of DNA ligase and plasmid vectors[J]. Nucleic Acids Research, 30 (2) : e2.
Kim JM, Kim JS, Park DH, et al. 2004. Improved recombinant gene expression in CHO cells using matrix attachment regions [J]. Biotechnol, 107 (2) : 95-105.
Kikuchi S, Satoh K, Nagata T, et al. 2003. Collection, mapping, and annotation of over 28,000 cDNA clones from japonica rice [J]. Science, 301: 376-379.
Kuo J, InmanJ, Brownstein M, et al. 2004. Evaluation of vector-primed cDNA library production from microgram quantities of total RNA [J]. Nucleic Acids Research, 32 (22) : el83.
Kato S, Ohtoko K, Ohtake H, et al. 2005. Vector-Capping: A Simple Method for Preparing a High-Quality Full-Length cDNA Library [J]. DNA Research, 12(1) : 53-62.
Karaba A, Dixit S, Greco R. 2007. Improvement of water use efficiency in rice by expression of HARDY, an Arabidopsis drought and salt tolerance gene [J]. Proc NatAcad Sci U. S. A. , 104. (39) : 15270-15275.
Li PH, Wang ZL, Zhang H, et al. 2004. Cloning and Expression Analysis of the B Subunit of V - H~+- ATPase in Leaves of Halophyte Suaeda salsa Under Salt Stress [J]. Acta Botanica Sinica, 46 (1) : 93-99.
Lawrence RJ, Pikaard CS. 2003. Transgene-induced RNA interference: a strategy for overcoming gene redundancy in polyploids to generate loos of-function mutations [J]. Plant, 36: 114-121.
Lin CW, Chang HB, Huang HJ. 2006. Zinc induces mitogen-activated protein kinase activation mediated by reactive oxygen species in rice roots[J]. Plant Physiol Biochem, 43 (10-11) : 963-968.
Lenassi M, Plemenitas A. 2007. Novel group VII histidine kinase HwHhk7B from the halophilic fungi Hortaea werneckii has a putative role in osmosensing [J]. Current Genetics, 51 (6) : 393-405.
Ma CL, Wang PP, Cao ZY, et al. 2002. cDNA Cloning and Gene Expression of APX in Suaeda salsa in Response to Salt Stress [J]. Journal of Plant Physiology and Molecular Biology, 28 (4) : 261-266.
Mehta PA, Sivaprakash K, Parani M, et al. 2005. Generation and analysis of expressed sequence tags from the salt-tolerant mangrove species Avicennia marina (Forsk) Vierh [J]. Theor Appl Genet, 110 (3) : 416-424.
Mayer H, Bauer H, Prohaska R. 2001. Organization and chromosomal localization of the human and mouse genes coding for LanC-like protein 1 (Lancl1) [J]. Cytogenet Cell Genetics, 93: 100-104.
Mankin SL, George C, Allen, et al. 2003. Elevation of transgene expression level by flanking matrix attachment regions (MAR) is promoter dependent: A study of the interactions of six promoters with the RB7 MAR [J]. Transgenic Research, (12) : 3-12.
Matzke MA, Mette MF, Matzke AJ. 2000. Transgene silencing by the host genome defense: implications for the evolution of epigenetic control mechanisms in plants and vertebrates [J]. Plant Mol Biol, 43 (223) : 401-415.
Moriwaki A, Kubo E, Arase S, Kihara J.2006. Disruption of SRM1, a mitogen-activated protein kinase gene , affects sensitivity to osmotic and ultraviolet stressors in the phytopathogenic fungus Bipolaris oryzae[J]. FEMS Microbiol Lett, 257 (2) : 253-61.
Motoyama T, Ohira T, Kadokura K, Ichiishi A, Fujimura M. Yamaguchi I, Kudo T . 2005 . An Os-1 family histidine kinase from a filamentous fungus confers fungicide-sensitivity to yeast[J]. Curr Genet, 47 (5) : 298-306.
Marie-Jo, Droillard, et al. 2002. Different Protein Kinase Families are Activated by Osmotic Stresses in Arabidopsis Thaliana Cell Suspensions: Involvement of the MAP Kinases AtMPK3 and AtMPK6[J]. FEBS Letters, 527: 43-50.
Morisawa G, Han-yama A, Moda I, et al. 2000. AHM1, A Novel Type of Nuclear Matrix-localized, MAR Binding Protein with a Single AT Hook and a Domain-homologous region[J]. Plant Cell, 12: 1903-1916.
Muskens MWM, Vissers APA, Mol JNM, et al. 2000. Role of Inverted DNA Repeats in Transcriptional and Posttranscriptional Gene Silencing[J]. Plant Mol Biol, 43: 243-260.
Ma SY, Wu WH. 2007. AtCPK 23 functions in Arabidopsis responses to drought and salt stresses [J]. Plant Molecular Biology, 65(4): 511-518.
Nowak W, Gawlowska M, Jarmolowski A, et al. 2001. Effect of Nuclear Matrix Attachment Regions on Transgene Expression in Tobacco Plants[J]. Acta Bioch Polonica, 48: 637-646.
Ning J, Yuan B, Xie KB, Hu HH, Wu CQ, Xiong LZ. 2006. Isolation and identification of SA and JA inducible protein kinase gene OsSJMK1 in rice[J]. Yi Chuan Xue Bao, 33 (7): 625-633.
Oh1 JH, Kim YS, Kim NS. 2003. An improved method for constructing a full length enriched cDNA library using small amounts of total RNA as a starting material [J]. Experimental and Molecular Medicine, 35 (6) : 586-590.
Oh DH, Gong QQ, Ulanov A, et al. 2007. Sodium stress in the halophyte Thellungiella halophila and transcriptional changes in a thsosl-RNA interference line[J]. Journal of Integrative Plant Biology, 49 (10) : 1484-1496.
Ortega D, Raynal M, Laudie M , et al. 2002. Flanking sequence tags in Arabidopsis thaliana T-DNA insertion lines: a pilot study[J]. C R Biol, 325 (7) : 773-780.
Peng YH, Lin WL, Cai WM, et al. 2007. Overexpression of a Panax ginseng tonoplast aquaporin alters salt tolerance, drought tolerance and cold acclimation ability in transgenic Arabidopsis plants [J]. Planta, 226 (3) : 729-740.
Paul R Muskett, Leah Clissold, Adriano Marocco, et al. 2003. A resource of mapped dissociation launch pads for targeted insertional mutagenesis[J]. Plant Physiology, 132: 506-516.
Qutob D, Hraber PT, Sobral BWS, et al. 2000. Comparative analysis of expressed sequences in Phytophthora sojae[J]. Plant Physiology, 123 (1) : 243-253.
Reyna NS, Yang Y. 2006. Molecular analysis of the rice MAP kinase gene family in relation to Magnaporthe grisea infection[J]. Mol Plant Microbe Interact, 19 (5) : 530-40.
Song D, Chen J, Song F, Zheng Z. 2006. A novel rice MAPK gene, OsBIMK2, is involved in disease-resistance responses[J]. Plant Biol (Stuttg), 8 (5) : 587-96.
SHEVCHU KNA, BRYKSIN AV, NUSINOVICH YA, et al. 2004. Construction of long DNA molecules using long PCR based fusion of several fragments simultaneously [J]. Nucleic Acids Research, 32 (2) : 19.
Shinji Kawasaki, Chris Borchert. 2001. Gene expression profiles during the initial phase of salt stress in rice[J]. Plant Cell, 13: 889-905.
Seki M, Narusaka M, Abe H, et al. 2001. Monitoring the expression pattern of 1300 Arabidopsis genes under rought and cold stresses by singe full length cDNA microarray [J]. Plant Cell, 13 (1) : 61-72.
Shinji Kawasaki, Chris Borchert. 2001. Gene Expression Profiles during the Initial Phase of Salt Stress in Rice[J]. Plant Cell, 13: 889-905.
Seki M, Narusaka M, Abe H, et al. 2001. Monitoring the Expression Pattern of 1300 Arabidopsis Genes under Rought and cold Stresses by Singe Full Length cDNA Microarray [J]. Plant Cell, 13 (1) : 61-72.
Sherry LeClere and Bonnie Bartel. 2001. A library of Arabidopsis 35S-cDNA lines for identifying novel mutants[J]. Plant Molecular Biology, 46: 695-703.
Shen W, Gomez Cadenas A, Routly E L, et al. 2001. The Salt Stress-Inducible Protein Kinase Gene, Esi47, from the Salt-Tolerant Wheatgrass Lophopyrum elongatum Is Involved in Plant Hormone Signaling [J]. Plant Physiology, 125 (3) : 1429-1441.
Shen YG, Y an DQ, Zhang W K, et al. 2003. Novel Halophyte EREBP/ AP2-type DNA Binding Protein Improves Salt Tolerance in Transgenic Tobacco [J]. Acta Botanica Sinica, 45(1): 82-87.
Sughura Y, Carninci P, Iton M, et al. 2001. Comparative evaluation of 5-end-sequence quality of clones in CAP trapper and other full length cDNA libraries [J]. Gene, 263:93-102.
Tague BW. 2001. Germ-line transformation of Arabidopsis lasiocarpa [J]. Transgenic Res, 10: 259-267.
Tikhonov AP, Bennetzen JL, Avramova ZV. 2000. Structural Domains and Matrix Attachment Regions Along Colinear Chromosomal Segments of Maize and Sorghum[J]. Plant Cell, 12: 249-264.
Tokihiko Nanjo, Norihiro Futamura, Mitsuru Nishiguchi. 2004. Characterization of Full-length Enriched Expressed Sequence Tags of Stress-treated Poplar Leaves[J]. Plant and Cell Physiology, 45 (12) : 1738-1748.
WM ichalek, WWeschke, KPP leissner, et al. 2002. EST Analysis in Barley Defines a Unigeneset Comprising 4000 Genes [J]. Theor Appl Genet, 104: 97-103.
Wei Ji, Yong Li, Jie Li, Cui-hong Dai, Xi Wang, Xi Bai, Hua Cai, Liang Yang and Yan-ming Zhu. 2006. Generation and analysis of expressed sequence tags from NaCl-treated Glycine soja. BMC Plant Biology, 6: 4.
Wang Hui wen, Zhang Jin song, Gai JunYi, et al. 2006. Cloning and comparative analysis of the gene encoding diacylglycerol acyltransferase from wild type and cultivated soybean [J]. Thero Appl Genet, 112 : 1086-1097.
Wang YC, Chu YG, Liu GF, et al. 2007. Identification of expressed sequence tags in an alkali grass ( Puccinellia tenuiflora) cDNA library [J]. Journal of Plant Physiology, 164 (1) : 78-89.
Wu YX, Ding N, Zhao X, et al. 2007. Molecular characterization of PeSOS1: the putative Na+/ H+ antiporter of Populus euphratica [J]. Plant Molecular Biology, 65 (1-2) : 1-11.
Wang ZL, Li PH, Fredricksen M, et al. 2004. Expressed sequence tags from Thellungiella halophila, a new model to study plant salt-tolerance [J]. Plant Science, 166 (3) : 609-616.
Wong CE, Li Y, Whitty BR, et al. 2005. Expressed sequence tags from the Yukon ecotype of Thellungiella reveal that gene expression in response to cold, drought and salinity shows little overlap [J]. Plant Molecular Biology, 58 (4) : 561-574.
Wellenreuther R, Schupp I, Poustka A, et al . 2004. SMART amplification combined with cDNA size fractionation in order to obtain large full-length clones [J]. BMC Genomics, 5(1): 36-43.
Wiemann S, Mehrle A, Bechtel S, et al. 2003. cDNAs for functional genomics and proteomics: the German Consortium[J]. C R Biology, 326 (10-11) : 1003-1009.
Wang TY, Hou WH, Chai YR, et al. 2005. Nuclear Matrices and Matrix Attachment Regions from Green Alga: Dunaliella salina [J]. Acta G enet Sin, 32 (12) : 1312-1318.
Wolfgang M, Schmid T, Manfred W, et al. 1999. CapSelect: A highly sensitive method for 5-CAP-dependent enrichment of full-length cDNA in PCR-mediated analysis of mRNAs[J]. Nucleic Acids Research, 27 (21) : e31.
Xu J, Li HD, Chen LQ, et al. 2006. A protein kinase, interacting with two calcineurin B-like proteins, regulates K+ transporter AKT1 in Arabidopsis [J]. Cell, 125 (7): 1347-1360.
Xun Gu. 2003. Evolution of duplicate genes versus genetic robustness against null mutations[J]. Trends In Genetics, 19 (7) : 354-356.
Zhumabayeva B, Chenchik A, Siebert PD. 1999. RecA-mediated affinity capture: a method for full-length cDNA cloning[J]. Biotechniques, 27 (4) : 834-840.
Zhulidov PA, Bogdanova EA, Shcheglov AS, et al. 2004. Simple cDNA normalization using kamchatka crab duplex-specific nuclease [J]. Nucleic Acids Research, 32 (3) : 37.
Zhu YY, Machleder EM, Chenchik A, et al. 2001. Reverse transcriptase template switching: a SMART approach for full-length cDNA library construction[J]. Biotechniques, 30 (4) : 892-897.
Zhang BC, Cao L, Qian J, et al. 2002. Construction of directional cDNA library from human embryo nasopharyngeal epithelia and screeming a candidate tumor suppress or gene related with nasopharyngeal carcinoma[J]. Progress in Biochemistry and Biophysics, 29(2): 302-306.
Zhulidov PA, Bogdanova EA, Shcheglov AS, et al. 2005. A method for the preparation of normalized cDNA libraries enriched with full-length sequences [J]. Russian Journal of Bioorganic Chmistry, 31 (2) : 170-177.
Zouari N, Saad RB, Legavre T, et al. 2007. Identification and sequencing of ESTs from the halophyte grass Aeluropus littoralis [J]. Gene, 404 (1-2) : 61-69.
Zhang L, Ma XL, Zhang Q, et al. 2001. Expressed sequence tags from a NaCl-treated Suaeda salsa cDNA library [J]. Gene, 267 (2) : 193-200.
Zhao FY, Wang ZL, Zhang Q, et al. 2006. Analysis of the physiological mechanism of salt-tolerant transgenic rice carrying a vacuolar Na+/H+antiporter gene from Suaeda salsa [J]. Journal of Plant Research, 119 (2) : 95-104.
Zeng-lan Wang, Ping-hua Li, Mark Fredricksen. 2004. Expressed sequence tags from Thellungiella halophila, a new model to study plant salt-tolerance [J]. Plant Science, 166: 609-616.
董艳珍,李关荣.2004.RNA干扰的研究进展[J].生物技术通讯,15(1):60-62.
代翠红,李杰,朱延明,纪巍,柏锡.2005.不同DNA提取方法对4种重要作物DNA提取效率的比较[J].东北农业大学学报,36(3):329-332.
傅作中,程远国.张玉静等.2002.用长距PCR法构建恶性疟原虫cDNA表达文库[J].热带医学杂志,2(3):225-229.
付绍红,牛应泽,杨洪全等.2004.表面活性剂silwetL-77对floral-dip转化甘蓝型油菜效果的影响[J].分子植物育种,2(5):661-666.
高健,许晓风,乌慧玲,等.2004.特异种质烟草全长cDNA文库的构建及质量分析[J].安徽农业大学学报,31(1):22-25.
关荣霞,刘秀敏,常汝镇等.2006.辽宁新宾县原位保护区野生大豆(Glycine soja Sieb.& Zucc.)遗传多样性分析[J].高技术通讯,16(1):67-72.
海林,王克晶.2002.半野生大豆种质资源SSR位点遗传多样性分析[J].西北植物学报,22(4):751-757.
何宝坤,王玉洁,孙立平.2005.蛋白质组学研究技术及其在植物抗渗透胁迫研究中的应用[J].生物技术通报,2:18-22.
纪巍,李杰,朱延明,柏锡,代翠红,侯爱菊.2005.不同启动子调控的DREB1A 基因对黄瓜的遗传转化[J].东北农业大学学报,36(4):442-447.
纪宗玲,刘继中,陈苏民.2002.基因功能的研究方法[J].生物工程学报,18:116-120.
来永才,林红,方万成等.2005.黑龙江省野生大豆优异资源筛选、评价及利用研究[J].中国农学通报,21(6):379-382.
李杰,李晶,朱延明,张晶,代翠红,纪巍,才华.2003.DREB1A基因植物表达载体的构建[J].东北农业大学学报,34(2):199-204.
李俊华,张艳春,徐云远等.2004.拟南芥室内培养技术[J].植物学通报,21(2):201-204.
李向华,王克晶,李福山等.2005.野生大豆(Glycine soja)研究现状与建议[J].大豆科学,24(4):305-309.
李向华,田子罡,李福山.2003.新考察收集野生大豆与已保存野生大豆的遗传多样性比较[J].植物遗传资源学报,4(4):345-349.
李秋莉,杨华等.2002.植物甜菜碱合成酶的分子生物学和基因工程[J].生物工程进展,22(1):84-86.
李旭刚,路子显,陈蕾等.2001.豌豆核基质结合区的分离及其在转基因烟草中的功能分析[J].中国科学(C辑),31(3):230-237.
梁慧敏,夏阳,王太明.2003.植物抗寒冻、抗旱、耐盐基因工程研究进展[J].草业学报,12(3):1-7.
刘西燕.2008.OsMAPK4基因水稻突变体的培育及功能分析.东北农业大学,43.
刘文华,王义良,陈慧萍等.2002.玫瑰红绿海葵触手cDNA表达文库的构建和初步分析[J].生物工程学报,18(6):749-753.
刘凡,王国英,曹鸣庆.2003.农杆菌介导的植物原位转基因方法研究进展[J].分子植物育种,1(1):108-116.
刘守伟,刘士勇.2007.拟南芥室内培养技术研究[J].东北农业大学学报,38(2):279-281.
刘峰,施卫明.2006.拟南芥室内水培方法的改进[J].土壤,38(1):102-105.
刘悦萍,宫飞,赵晓萌.2005.水杨酸介导的信号转导途径与植物抗逆性[J].中国农学通报,21(7):227-229.
罗会颖,姚斌,袁铁铮等.2004.来源于Escherichia coli的高比活植酸酶基因的高效表达[J].生物工程学报,20(1):78-84.
吕宪禹,卢茜,刘桂琴,等.2002.导入野生大豆DNA小麦后代的农艺性状研究[J].南开大学学报:自然科学版,35(4):103-105.
马艺沔.2003.利用转基因拟南芥研究质外体CaM在植物生长发育中的作用[D].毛新国,孔秀英,赵光耀等.2005.利用改进的Cap-trapper法构建拟斯卑尔脱山羊草(Ae.speltoides Tausch)全长cDNA文库[J].遗传学报,32(8):811-817.
萨姆布鲁克,拉塞尔.2002.分子克隆实验指南[M].黄培堂等译.科学出版社.
苏金,朱汝财.2001.渗透胁迫调节的转基因表达对植物抗旱耐盐性的影响[J].植物学通报,18(2):129-136.
陶爱林,林兴华,张端品.2003.获取全长cDNA若干方法的比较[J].武汉植物学研究,21(2):179-186.
田佳.2008.拟南芥渗透胁迫应答基因的筛选及功能分析.东北农业大学,42.
涂艳阳,徐如祥,杨志林等.2003.人脑瘤组织全长cDNA文库的构建[J].第一军医大学学报,23(9):916-920.
王秀玲,卢茜,刘君等.2003.野生大豆DNA导入小麦及RAPD分子验证[J].南开大学学报:自然科学版,36(2):37-40.
王皓,康现江,王琦.2007.重组PCR技术研究进展和应用[J].中国生物工程杂志,27(5):153-156.
王曙明,孙寰,王跃强等.2002.大豆杂种优势及其高优势组合选配的研究:F1代籽粒产量的杂种优势与高优势组合选配[J].大豆科学,21(3):161-167.
王敏,刘萍,石明旺等.2005.野生大豆种子cDNA文库的构建与分析[J].生物技术通讯,16(5):509-511.
王爱荣,刘丽,周洁等.2005.拟南芥室內简易栽培和田间大规模种植的方法[J].福建农林大学学报:自然科学版,34(2):252-254.
王克晶,李福山.2000.我国野生大豆(Glsoja)种质资源及其种质创新利用[J].中 国农业科技导报,2(6).69-72.
王关林,方宏筠.2002.植物基因工程原理与技术[M].北京:科学出版社,473-474.
王天云,柴玉荣,袁保梅等.2004.核基质结合区对转基因表达的影响及其作用机制[J].细胞生物学杂志,26(6):587-590.
王艺磊,张子平.2003.日本对虾精巢和卵巢全长cDNA文库的构建[J].动物学杂志,38(2):9-13.
吴才君,范涉英.2004.植物转基因沉默[J].江西农业大学学报:155-158.
徐光硕,饶勇强,陈雁等.2004.用in planta方法转化甘蓝型油菜[J].作物学报,1:1-5.
徐春晖,夏光敏,贺晨霞.2002.农杆菌转化系统研究进展[J].生命科学,4(4):224-225.
许亮,卢向阳,田云.2003.后基因组时代基因功能分析的策略[J].中国生物工程杂志,23:29-34.
许志茹,李玉花.2004.基因芯片技术在植物研究中的应用[J].生物技术,14(6):70-72.
杨洪强,梁小娥.2003.高等植物中的CDPK及其生理生化功能[J].山东农业大学学报(自然科学版),34(2):284-288.
杨光宇,王洋,马晓萍等.2005.野生大豆种质资源评价与利用研究进展[J].吉林农业科学,30(2):61-63.
袁盛凌,段海清,张兆山.2003.蛇类凝血酶Calobin cDNA的设计合成与克隆[J].生物技术通讯,14(5):357-360.
于玲,王莱,牛吉山,陈佩度.2003.植物功能基因组研究进展[J].西北师范大学学报,39:104-116.
张可伟,王健美,郑成超.2004.核基质结合序列(MAR)与基因表达调控[J].生物工程学报,20:6-9.
张震祯,徐煜泉,黄海.2006.拟南芥-一把通往生命科学奥秘大门的钥匙[J].生命科学,18(5):442-446.
赵洪锟,王玉民,李启云等.2001.中国不同纬度野生大豆和栽培大豆SSR分析[J].大豆科学,20(3):172-176.
赵团结,盖钧镒.2006.大豆不育细胞质资源的发掘与鉴定[J].作物学报,32(11):1604-1610.
周晓馥,庄炳昌,王玉民等.2002.利用RAPD与SSR技术进行野生大豆种群内分化的研究[J].中国生态农业学报,10(4):629.
庄炳昌.1999.中国野生大豆研究二十年[J].吉林农业科学,24(5):3-10.
朱维岳,周桃英,钟明等.2006.基于遗传多样性和空间遗传结构的野生大豆居群采样策略[J].复旦大学学报:自然科学版,45(3):321-327.
Allen GC,Spiker S,Thompson WF.2000.Use of matrix attachment regions(MARs)to minimize transgene silencing[J].Plant Mol Biol,43(223):361-376.
Ahre n D, Tholander M, Fekete C, et al. 2005. Comparison of gene expression in trap cells and vegetative hyphae of the nematophag ous fungus Monacrosporium haptotylum [J]. Microbiology, 51 (3) : 789-803.
Brouwer C, Bruce W, Maddock S, et al. 2003. Suppression of transgene Silencing by Matrix Attachment Regions in Maize. A dual Role for the Maize 5'-ADHl Matrix Attachment Region[J]. Plant Cell, 14: 2251-2264.
Butaye KM, Goderis IJ, Wouters PF, et al. 2004. Stable high-level transgene expression in Arabidopsis thaliana using gene silencing mutants and matrix attachment regions [J]. Plant J, 39 (3) : 440-449.
Carninci P, Waki K, Shiraki T, et al. 2003 . Targeting a complex transcriptome: the construction of the mouse full-length cDNA encyclopedia [J]. Genome Research, 13: 1273-1289.
Chenchik A, Moqadam F, Siebert P. 1996. RNA: Isolation, Analysis and Synthesis [M]. New York: Wiley-Liss, 273-321.
Curtis IS, Nam HG. 2001. Transgenic radish (Raphanus sativus L. Iongipinnatus Bailey) by floral-dip method plant development and surfactant are important in optimizing transformation efficiency [J]. Transgenic Research, 10: 363-371.
Curtis IS. 2005. Production of transgenic crops by the floral-dip method[J]. Methods Mol Biol, 286: 103-106.
Clough SJ. 2005. Floral dip: agrobacterium-mediated germ line transformation [J]. Methods Mol Biol, 286: 91-102.
Cheng YX, Long M. 2007. A cytosolic NADP-malic enzyme gene from rice ( Oryza sativa L. ) confers salt tolerance in transgenic Arabidopsis [J]. Biotechnology Letters, 29 (7) : 1129-1134.
Chen M, Wang QY, Cheng XG, et al. 2007. GmDREB2, a soybean DRE-binding transcription factor , conferred drought and high-salt tolerance in transgenic plants [J]. Biochemical and Biophysical Research Communications, 353 (2) : 299-305.
Chen SH, Guo SL, Wang ZL, et al. 2007. Expressed sequence tags from the halophyte Limonium sinense [J]. DNA Sequence, 18 (1) : 61-67.
Dastidar K G, Maitra S, Goswami L, et al. 2006. An insight into the molecular basis of salt tolerance of L-my-oinositol 1 -Psynthase (PcINO 1) from Porteresia coarctata (Roxb. ) Tateoka, a halophytic wild rice [J]. Plant Phsiology, 140 (4) : 1279-1296.
Francois Chaumont, Menachem Moshelion, Mark J, Danielst. 2005. Regulation of plant aquaporin activity[J]. Biol Cell, 97: 749-764.
Francois T, Anne P, Pierre B, et al. 2005 . Enhancement of maize transformation efficiency by the use of maize matrix attachment regions [J]. Plant Cell, 80: 295-303.
Fu SF, Chou WC, Huang DD, Huang HJ. 2002. Transcriptional regulation of a rice mitogen-activated protein kinase gene , OsMAPK , in response to environmental stresses[J]. Plant Cell Physiol, 43 (8) : 958-63.
Fu XH, Huang YL, Deng SL, et al. 2005. Construction of a SSH library of Aegiceras corniculatum under salt stress and expression analysis of four transcripts [J]. Plant Science, 169 (1) : 147-154.
Glazko G V, Rogozin I B, Glazkov M V. 2001. Comparative Study and Prediction of DNA Fragments Associated with Various Element of the Nuclear Matrix [J]. Biochem BiophysActa, 1517: 351-364.
Gernot G. 2003. Mapping multiple consequenced T-DNA integration sites within the Arabidopsis genome[J]. Bioinformatics, 19 (5) : 579-586.
Guo XL, Cao YR, Cao ZY, et al. 2004. Molecular cloning and characterization of a stress-induced peroxiredoxin Q gene in halophyte Suaedasalsa [J]. Plant Science, 167(5) : 969-975.
Gao F, Gao Q, Duan X G, et al. 2006. Cloning of an H~+-PPase gene from Thellungiella halophila and its heterologous expression to improve tobacco salt tolerance [J]. Journal of Eeperimental Botany, 57 (12) : 3259-3270.
Guo SL, Yin HB, Zhang X, et al . 2006. Molecular cloning and characterization of a vacuolarn H~+-pyrophosphatase gene, SsVP, from the halophyte Suaeda salsa and its over-expression increases salt and drought tolerance of Arabidopsis [J]. Plant Molecular Biology, 60 (1) : 41-50.
Geng DG, Han Y, Wang YQ, et al. 2004. Construction of a system for the stable expression of foreign genes in Dunaliella salina [J]. Acta Bot Sin, 46 (3) : 342-346.
Girod PA, Zahn Zabal M, Mermod N. 2005. Use of the chicken lysozyme 5'matrix attachment region to generate high producer CHO cell lines [J]. Biotechnol Bioeng, 91 (1) : 121.
Gspar YM. 2004. Characterizati on of the Arabidopsis lysine-rich arabinogalactan protein AtAGP17 mutant (rat1) that results in a decreased efficiency of agrobacterium transformation [J]. Plant Physiol, 135 (4) : 2162-71.
Garcia Mata C. Lamattina L. 2001. Nitric oxide induces stomatal closure and enhances the adaptive plant response against drought stress[J]. Plant Physiol, 12 (6) : 1196-1204.
Hu HH, Dai MQ, Yao JL, et al. 2006. Over-expressing a NAM, ATAF, and CUC (NAC) transcription factor enhances drought resistance and salt tolerance in rice [J]. Proc Nat Acad Sci U. S. A. , 103 (35) : 12987-12992.
Hmida-Sayari A, Gargouri-Bouzid R, Bidani A, et al. 2005. Over-expression of Delta ( 1 ) - pyrroline-5-carboxylate synthetase increases praline production and confers salt tolerance in transgenic potato plants [J]. Plant Science, 169 (4) : 746-752. Halweg C, Thompson WF, Spiker S. 2005. The rb7 matrix attachment region increases the likelihood and magnitude of transgene expression in tobacco cells: a flow cytometric study [J]. Plant Cell, 17 (2) : 418-429.
Ichikawa T, Nakazawa M, Kawashima M, el al. 2003. Sequence database of 1172 T-DNA insertion sites in Arabidopsis activation-tagging lines that showed phenotypes in T1 generation[J]. Plant J, 36 (3) : 421-429.
Jiang GZ, Lu YM, Niu XL, et al. 2005. The actin gene promoter driven bar as a dominant selectable marker for nuclear trans formation of Dunaliella salina [J]. Acta Genet Sin, 32 (4) : 424-433.
JiaJ, FuJ, Zheng J, et al. 2006. Annotation and expression profile analysis of 2073 full-length cDNAs from stress-induced maize (Zea mays L. ) seedlings [J]. Plant J, 48 (5) : 710-727.
KY amaguchi, Shinozaki K. 2003. OsDREB genes in rice, Oryza sativa L, encode transcription activators that function in drought, high-salt and cold responsive gene expression[J]. Plant J, (4) : 751-763.
Kurusu T, Yagala T, Miyao A, Hirochika H, Kuchitsu K. 2005. Identification of a putative voltage-gated Ca~(2+) channel as a key regulator of elicitor-induced hypersensitive cell death and mitogen-activated protein kinase activation in rice[J]. Plant, 42 (6) : 798-809.
KUWAYAMA H, OBARA S, MORIO T, et al. 2002. PCR-mediated generation of a gene disruption construct without the use of DNA ligase and plasmid vectors[J]. Nucleic Acids Research, 30 (2) : e2.
Kim JM, Kim JS, Park DH, et al. 2004. Improved recombinant gene expression in CHO cells using matrix attachment regions [J]. Biotechnol, 107 (2) : 95-105.
Kikuchi S, Satoh K, Nagata T, et al. 2003. Collection, mapping, and annotation of over 28,000 cDNA clones from japonica rice [J]. Science, 301: 376-379.
Kuo J, InmanJ, Brownstein M, et al. 2004. Evaluation of vector-primed cDNA library production from microgram quantities of total RNA [J]. Nucleic Acids Research, 32 (22) : el83.
Kato S, Ohtoko K, Ohtake H, et al. 2005. Vector-Capping: A Simple Method for Preparing a High-Quality Full-Length cDNA Library [J]. DNA Research, 12(1) : 53-62.
Karaba A, Dixit S, Greco R. 2007. Improvement of water use efficiency in rice by expression of HARDY, an Arabidopsis drought and salt tolerance gene [J]. Proc NatAcad Sci U. S. A. , 104. (39) : 15270-15275.
Li PH, Wang ZL, Zhang H, et al. 2004. Cloning and Expression Analysis of the B Subunit of V - H~+- ATPase in Leaves of Halophyte Suaeda salsa Under Salt Stress [J]. Acta Botanica Sinica, 46 (1) : 93-99.
Lawrence RJ, Pikaard CS. 2003. Transgene-induced RNA interference: a strategy for overcoming gene redundancy in polyploids to generate loos of-function mutations [J]. Plant, 36: 114-121.
Lin CW, Chang HB, Huang HJ. 2006. Zinc induces mitogen-activated protein kinase activation mediated by reactive oxygen species in rice roots[J]. Plant Physiol Biochem, 43 (10-11) : 963-968.
Lenassi M, Plemenitas A. 2007. Novel group VII histidine kinase HwHhk7B from the halophilic fungi Hortaea werneckii has a putative role in osmosensing [J]. Current Genetics, 51 (6) : 393-405.
Ma CL, Wang PP, Cao ZY, et al. 2002. cDNA Cloning and Gene Expression of APX in Suaeda salsa in Response to Salt Stress [J]. Journal of Plant Physiology and Molecular Biology, 28 (4) : 261-266.
Mehta PA, Sivaprakash K, Parani M, et al. 2005. Generation and analysis of expressed sequence tags from the salt-tolerant mangrove species Avicennia marina (Forsk) Vierh [J]. Theor Appl Genet, 110 (3) : 416-424.
Mayer H, Bauer H, Prohaska R. 2001. Organization and chromosomal localization of the human and mouse genes coding for LanC-like protein 1 (Lancl1) [J]. Cytogenet Cell Genetics, 93: 100-104.
Mankin SL, George C, Allen, et al. 2003. Elevation of transgene expression level by flanking matrix attachment regions (MAR) is promoter dependent: A study of the interactions of six promoters with the RB7 MAR [J]. Transgenic Research, (12) : 3-12.
Matzke MA, Mette MF, Matzke AJ. 2000. Transgene silencing by the host genome defense: implications for the evolution of epigenetic control mechanisms in plants and vertebrates [J]. Plant Mol Biol, 43 (223) : 401-415.
Moriwaki A, Kubo E, Arase S, Kihara J.2006. Disruption of SRM1, a mitogen-activated protein kinase gene , affects sensitivity to osmotic and ultraviolet stressors in the phytopathogenic fungus Bipolaris oryzae[J]. FEMS Microbiol Lett, 257 (2) : 253-61.
Motoyama T, Ohira T, Kadokura K, Ichiishi A, Fujimura M. Yamaguchi I, Kudo T . 2005 . An Os-1 family histidine kinase from a filamentous fungus confers fungicide-sensitivity to yeast[J]. Curr Genet, 47 (5) : 298-306.
Marie-Jo, Droillard, et al. 2002. Different Protein Kinase Families are Activated by Osmotic Stresses in Arabidopsis Thaliana Cell Suspensions: Involvement of the MAP Kinases AtMPK3 and AtMPK6[J]. FEBS Letters, 527: 43-50.
Morisawa G, Han-yama A, Moda I, et al. 2000. AHM1, A Novel Type of Nuclear Matrix-localized, MAR Binding Protein with a Single AT Hook and a Domain-homologous region[J]. Plant Cell, 12: 1903-1916.
Muskens MWM, Vissers APA, Mol JNM, et al. 2000. Role of Inverted DNA Repeats in Transcriptional and Posttranscriptional Gene Silencing[J]. Plant Mol Biol, 43: 243-260.
Ma SY, Wu WH. 2007. AtCPK 23 functions in Arabidopsis responses to drought and salt stresses [J]. Plant Molecular Biology, 65(4): 511-518.
Nowak W, Gawlowska M, Jarmolowski A, et al. 2001. Effect of Nuclear Matrix Attachment Regions on Transgene Expression in Tobacco Plants[J]. Acta Bioch Polonica, 48: 637-646.
Ning J, Yuan B, Xie KB, Hu HH, Wu CQ, Xiong LZ. 2006. Isolation and identification of SA and JA inducible protein kinase gene OsSJMK1 in rice[J]. Yi Chuan Xue Bao, 33 (7): 625-633.
Oh1 JH, Kim YS, Kim NS. 2003. An improved method for constructing a full length enriched cDNA library using small amounts of total RNA as a starting material [J]. Experimental and Molecular Medicine, 35 (6) : 586-590.
Oh DH, Gong QQ, Ulanov A, et al. 2007. Sodium stress in the halophyte Thellungiella halophila and transcriptional changes in a thsosl-RNA interference line[J]. Journal of Integrative Plant Biology, 49 (10) : 1484-1496.
Ortega D, Raynal M, Laudie M , et al. 2002. Flanking sequence tags in Arabidopsis thaliana T-DNA insertion lines: a pilot study[J]. C R Biol, 325 (7) : 773-780.
Peng YH, Lin WL, Cai WM, et al. 2007. Overexpression of a Panax ginseng tonoplast aquaporin alters salt tolerance, drought tolerance and cold acclimation ability in transgenic Arabidopsis plants [J]. Planta, 226 (3) : 729-740.
Paul R Muskett, Leah Clissold, Adriano Marocco, et al. 2003. A resource of mapped dissociation launch pads for targeted insertional mutagenesis[J]. Plant Physiology, 132: 506-516.
Qutob D, Hraber PT, Sobral BWS, et al. 2000. Comparative analysis of expressed sequences in Phytophthora sojae[J]. Plant Physiology, 123 (1) : 243-253.
Reyna NS, Yang Y. 2006. Molecular analysis of the rice MAP kinase gene family in relation to Magnaporthe grisea infection[J]. Mol Plant Microbe Interact, 19 (5) : 530-40.
Song D, Chen J, Song F, Zheng Z. 2006. A novel rice MAPK gene, OsBIMK2, is involved in disease-resistance responses[J]. Plant Biol (Stuttg), 8 (5) : 587-96.
SHEVCHU KNA, BRYKSIN AV, NUSINOVICH YA, et al. 2004. Construction of long DNA molecules using long PCR based fusion of several fragments simultaneously [J]. Nucleic Acids Research, 32 (2) : 19.
Shinji Kawasaki, Chris Borchert. 2001. Gene expression profiles during the initial phase of salt stress in rice[J]. Plant Cell, 13: 889-905.
Seki M, Narusaka M, Abe H, et al. 2001. Monitoring the expression pattern of 1300 Arabidopsis genes under rought and cold stresses by singe full length cDNA microarray [J]. Plant Cell, 13 (1) : 61-72.
Shinji Kawasaki, Chris Borchert. 2001. Gene Expression Profiles during the Initial Phase of Salt Stress in Rice[J]. Plant Cell, 13: 889-905.
Seki M, Narusaka M, Abe H, et al. 2001. Monitoring the Expression Pattern of 1300 Arabidopsis Genes under Rought and cold Stresses by Singe Full Length cDNA Microarray [J]. Plant Cell, 13 (1) : 61-72.
Sherry LeClere and Bonnie Bartel. 2001. A library of Arabidopsis 35S-cDNA lines for identifying novel mutants[J]. Plant Molecular Biology, 46: 695-703.
Shen W, Gomez Cadenas A, Routly E L, et al. 2001. The Salt Stress-Inducible Protein Kinase Gene, Esi47, from the Salt-Tolerant Wheatgrass Lophopyrum elongatum Is Involved in Plant Hormone Signaling [J]. Plant Physiology, 125 (3) : 1429-1441.
Shen YG, Y an DQ, Zhang W K, et al. 2003. Novel Halophyte EREBP/ AP2-type DNA Binding Protein Improves Salt Tolerance in Transgenic Tobacco [J]. Acta Botanica Sinica, 45(1): 82-87.
Sughura Y, Carninci P, Iton M, et al. 2001. Comparative evaluation of 5-end-sequence quality of clones in CAP trapper and other full length cDNA libraries [J]. Gene, 263:93-102.
Tague BW. 2001. Germ-line transformation of Arabidopsis lasiocarpa [J]. Transgenic Res, 10: 259-267.
Tikhonov AP, Bennetzen JL, Avramova ZV. 2000. Structural Domains and Matrix Attachment Regions Along Colinear Chromosomal Segments of Maize and Sorghum[J]. Plant Cell, 12: 249-264.
Tokihiko Nanjo, Norihiro Futamura, Mitsuru Nishiguchi. 2004. Characterization of Full-length Enriched Expressed Sequence Tags of Stress-treated Poplar Leaves[J]. Plant and Cell Physiology, 45 (12) : 1738-1748.
WM ichalek, WWeschke, KPP leissner, et al. 2002. EST Analysis in Barley Defines a Unigeneset Comprising 4000 Genes [J]. Theor Appl Genet, 104: 97-103.
Wei Ji, Yong Li, Jie Li, Cui-hong Dai, Xi Wang, Xi Bai, Hua Cai, Liang Yang and Yan-ming Zhu. 2006. Generation and analysis of expressed sequence tags from NaCl-treated Glycine soja. BMC Plant Biology, 6: 4.
Wang Hui wen, Zhang Jin song, Gai JunYi, et al. 2006. Cloning and comparative analysis of the gene encoding diacylglycerol acyltransferase from wild type and cultivated soybean [J]. Thero Appl Genet, 112 : 1086-1097.
Wang YC, Chu YG, Liu GF, et al. 2007. Identification of expressed sequence tags in an alkali grass ( Puccinellia tenuiflora) cDNA library [J]. Journal of Plant Physiology, 164 (1) : 78-89.
Wu YX, Ding N, Zhao X, et al. 2007. Molecular characterization of PeSOS1: the putative Na+/ H+ antiporter of Populus euphratica [J]. Plant Molecular Biology, 65 (1-2) : 1-11.
Wang ZL, Li PH, Fredricksen M, et al. 2004. Expressed sequence tags from Thellungiella halophila, a new model to study plant salt-tolerance [J]. Plant Science, 166 (3) : 609-616.
Wong CE, Li Y, Whitty BR, et al. 2005. Expressed sequence tags from the Yukon ecotype of Thellungiella reveal that gene expression in response to cold, drought and salinity shows little overlap [J]. Plant Molecular Biology, 58 (4) : 561-574.
Wellenreuther R, Schupp I, Poustka A, et al . 2004. SMART amplification combined with cDNA size fractionation in order to obtain large full-length clones [J]. BMC Genomics, 5(1): 36-43.
Wiemann S, Mehrle A, Bechtel S, et al. 2003. cDNAs for functional genomics and proteomics: the German Consortium[J]. C R Biology, 326 (10-11) : 1003-1009.
Wang TY, Hou WH, Chai YR, et al. 2005. Nuclear Matrices and Matrix Attachment Regions from Green Alga: Dunaliella salina [J]. Acta G enet Sin, 32 (12) : 1312-1318.
Wolfgang M, Schmid T, Manfred W, et al. 1999. CapSelect: A highly sensitive method for 5-CAP-dependent enrichment of full-length cDNA in PCR-mediated analysis of mRNAs[J]. Nucleic Acids Research, 27 (21) : e31.
Xu J, Li HD, Chen LQ, et al. 2006. A protein kinase, interacting with two calcineurin B-like proteins, regulates K+ transporter AKT1 in Arabidopsis [J]. Cell, 125 (7): 1347-1360.
Xun Gu. 2003. Evolution of duplicate genes versus genetic robustness against null mutations[J]. Trends In Genetics, 19 (7) : 354-356.
Zhumabayeva B, Chenchik A, Siebert PD. 1999. RecA-mediated affinity capture: a method for full-length cDNA cloning[J]. Biotechniques, 27 (4) : 834-840.
Zhulidov PA, Bogdanova EA, Shcheglov AS, et al. 2004. Simple cDNA normalization using kamchatka crab duplex-specific nuclease [J]. Nucleic Acids Research, 32 (3) : 37.
Zhu YY, Machleder EM, Chenchik A, et al. 2001. Reverse transcriptase template switching: a SMART approach for full-length cDNA library construction[J]. Biotechniques, 30 (4) : 892-897.
Zhang BC, Cao L, Qian J, et al. 2002. Construction of directional cDNA library from human embryo nasopharyngeal epithelia and screeming a candidate tumor suppress or gene related with nasopharyngeal carcinoma[J]. Progress in Biochemistry and Biophysics, 29(2): 302-306.
Zhulidov PA, Bogdanova EA, Shcheglov AS, et al. 2005. A method for the preparation of normalized cDNA libraries enriched with full-length sequences [J]. Russian Journal of Bioorganic Chmistry, 31 (2) : 170-177.
Zouari N, Saad RB, Legavre T, et al. 2007. Identification and sequencing of ESTs from the halophyte grass Aeluropus littoralis [J]. Gene, 404 (1-2) : 61-69.
Zhang L, Ma XL, Zhang Q, et al. 2001. Expressed sequence tags from a NaCl-treated Suaeda salsa cDNA library [J]. Gene, 267 (2) : 193-200.
Zhao FY, Wang ZL, Zhang Q, et al. 2006. Analysis of the physiological mechanism of salt-tolerant transgenic rice carrying a vacuolar Na+/H+antiporter gene from Suaeda salsa [J]. Journal of Plant Research, 119 (2) : 95-104.
Zeng-lan Wang, Ping-hua Li, Mark Fredricksen. 2004. Expressed sequence tags from Thellungiella halophila, a new model to study plant salt-tolerance [J]. Plant Science, 166: 609-616.