新疆野生油菜BAC文库的构建及单条染色体克隆筛选的研究
|
摘要
油菜是重要的油料作物之一。新疆野生油菜是野芥(Sinapis arvensis)分布在中国新疆的一个自然群体,它生长势强,抗病虫害、抗菌核病能力强,具有不育胞质,是油菜育种的优良野生种质资源。甘蓝型油菜-新疆野生油菜二体异附加系(B. napus-S. arvensis disomic alien addition line)是新疆野生油菜18号与甘蓝型油菜(Brassica napus)品种中双4号原生质体融合,连续4次回交后得到的新型油菜恢复系,它具有一对来源于新疆野生油菜的染色体,该染色体上携带育性恢复基因,克隆该恢复基因对探索油菜育性恢复机制、培育优良转基因油菜恢复系具有重要的科学意义和生产价值。本研究构建了新疆野生油菜基因组BAC文库,并以甘蓝型油菜-新疆野生油菜二体异附加系BAC文库为材料,对筛选新疆野生油菜染色体来源的BAC克隆进行了研究。研究结果如下：
新疆野生油菜基因组BAC文库的构建：用低熔点琼脂糖包埋细胞核的方法制备高分子量DNA,HindⅢ酶切,电洗脱回收100-300kb大片段DNA,与pCC1BACHTM载体连接,用0.025μm的透析膜悬浮在10%的PEGS000中冰上脱盐浓缩,转化E.coilEP1300感受态细胞,得到新疆野生油菜基因组BAC文库,保存在38块384孔板中。随机挑取100个克隆,用Notl酶切,脉冲电泳检测插入片段大小,插入片段在50-250kb之间,平均插入片段105kb,该文库共有14592个克隆,覆盖了新疆野生油菜基因组4.3倍。
野生油菜特定染色体来源BAC克隆的筛选：以新疆野生油菜基因组DNA为探针,直接对甘蓝型油菜-新疆野生油菜二体异附加系BAC文库的克隆进行菌落原位杂交,以期获得野生油菜特定染色体来源的BAC克隆,但未获得理想的结果,下一步需要以合适剂量的甘蓝型油菜基因组DNA进行封阻等方法继续开展相关研究。
Rape is one of the important oil crops. Xinjiang wild rape is a wild mustard (Sinapis arvensis) found in a natural population of Xinjiang, China, it has strong growth potential, strong resistance to pests and diseases, and strong resistance to Sclerotinia sclerotiorum, as well as sterile cytoplasm, so it is a fine breeding of wild rapeseed germplasm. Brassica-Xinjiang wild rape disomic addition lines (B. napus-S. arvensis disomic alien addition line) is a new rapeseed restorer line which is obtained through protoplast fusion between Xinjiang wild rape 18 and ZhongShuang 4 (Brassica napus).and continuously backcross four times.The disomic addition lines have a pair of chromosomes from Xinjiang wild rape, which carrying the fertility restorer gene. Cloning of the restorer gene is to explore recovery mechanism, and cultivate good transgenic rapeseed restorer lines, It has important scientific significance and production value. The study has constructed wild rape genomic BAC library and studied BAC clones of Xinjiang wild rape chromosome from Brassica napus-Xinjiang wild rape disomic addition lines of BAC library. The results are as follows:
Construction of Xinjiang wild rape genomic BAC library:With low melting point agarose embedded nuclei,high molecular weight DNA were prepared, then digested by HindⅢ, through electro elution,100-300kb large fragment DNA were recovered and linked with pCC1BACHTM vector, the connection was desalted and concentrated with 0.025μm dialysis membrane suspended in 10% PEG8000 on ice, then transfered into E.coil EP1300 competent cells. Xinjiang wild rape genomic BAC library had been constructed, and stored in 38 384 well plates. Randomly selected 100 clones, digested with NotI, tested insert size with PFGE, the library has 14,592 clones, with the insert size of 50-250kb, average insert of 105kb, covering the genome of Xinjiang wild rape 4.3 times.
Screening specific BAC clones from wild rape chromosome:Xinjiang wild rape genomic DNA as a probe, directly hybridized colony in situ to the Brassica napus-Xinjiang wild rape disomic addition lines of clone BAC library to obtain a specific chromosome of wild rape sources of BAC clones, but get no desired results, the right dose of Brassica napus genome DNA for blocking will be carried on for the further correlation research.
新疆野生油菜基因组BAC文库的构建：用低熔点琼脂糖包埋细胞核的方法制备高分子量DNA,HindⅢ酶切,电洗脱回收100-300kb大片段DNA,与pCC1BACHTM载体连接,用0.025μm的透析膜悬浮在10%的PEGS000中冰上脱盐浓缩,转化E.coilEP1300感受态细胞,得到新疆野生油菜基因组BAC文库,保存在38块384孔板中。随机挑取100个克隆,用Notl酶切,脉冲电泳检测插入片段大小,插入片段在50-250kb之间,平均插入片段105kb,该文库共有14592个克隆,覆盖了新疆野生油菜基因组4.3倍。
野生油菜特定染色体来源BAC克隆的筛选：以新疆野生油菜基因组DNA为探针,直接对甘蓝型油菜-新疆野生油菜二体异附加系BAC文库的克隆进行菌落原位杂交,以期获得野生油菜特定染色体来源的BAC克隆,但未获得理想的结果,下一步需要以合适剂量的甘蓝型油菜基因组DNA进行封阻等方法继续开展相关研究。
Rape is one of the important oil crops. Xinjiang wild rape is a wild mustard (Sinapis arvensis) found in a natural population of Xinjiang, China, it has strong growth potential, strong resistance to pests and diseases, and strong resistance to Sclerotinia sclerotiorum, as well as sterile cytoplasm, so it is a fine breeding of wild rapeseed germplasm. Brassica-Xinjiang wild rape disomic addition lines (B. napus-S. arvensis disomic alien addition line) is a new rapeseed restorer line which is obtained through protoplast fusion between Xinjiang wild rape 18 and ZhongShuang 4 (Brassica napus).and continuously backcross four times.The disomic addition lines have a pair of chromosomes from Xinjiang wild rape, which carrying the fertility restorer gene. Cloning of the restorer gene is to explore recovery mechanism, and cultivate good transgenic rapeseed restorer lines, It has important scientific significance and production value. The study has constructed wild rape genomic BAC library and studied BAC clones of Xinjiang wild rape chromosome from Brassica napus-Xinjiang wild rape disomic addition lines of BAC library. The results are as follows:
Construction of Xinjiang wild rape genomic BAC library:With low melting point agarose embedded nuclei,high molecular weight DNA were prepared, then digested by HindⅢ, through electro elution,100-300kb large fragment DNA were recovered and linked with pCC1BACHTM vector, the connection was desalted and concentrated with 0.025μm dialysis membrane suspended in 10% PEG8000 on ice, then transfered into E.coil EP1300 competent cells. Xinjiang wild rape genomic BAC library had been constructed, and stored in 38 384 well plates. Randomly selected 100 clones, digested with NotI, tested insert size with PFGE, the library has 14,592 clones, with the insert size of 50-250kb, average insert of 105kb, covering the genome of Xinjiang wild rape 4.3 times.
Screening specific BAC clones from wild rape chromosome:Xinjiang wild rape genomic DNA as a probe, directly hybridized colony in situ to the Brassica napus-Xinjiang wild rape disomic addition lines of clone BAC library to obtain a specific chromosome of wild rape sources of BAC clones, but get no desired results, the right dose of Brassica napus genome DNA for blocking will be carried on for the further correlation research.
引文
1.薄存沛,闫晓红,徐有明,等.新疆野生油菜BAC文库的构建.中国油料作物学,2010,32(1)：14-19
2.陈庆祥.一种2n=18的芥菜型野生油菜.新疆农业科学,1986,1：18-19
3.陈书元,晁金泉,徐有明,等.甘蓝型油菜—新疆野生油菜二体异附加系BAC文库的构建.中国油料作物学报,2008,30(2)：137-142
4.程计华,李云昌,胡琼,等.油菜野芥NSa细胞质雄性不育系的特异性分子鉴定.作物学报,2008,34(11)：1946-1952
5.杜伟,王兆木,孙卫琳,等.新疆野生油菜体细胞染色体核型的研究.西北农业学报,1992,1(2)：8-10
6.杜伟,王兆木,孙卫琳,等.新疆野生油菜体细胞染色体数观察及统计.新疆农业科学,1992,4：156-157
7.官春云.新疆野生油菜与野芥(Sinapis arvensis L.)遗传性状的比较研究.作物学报,1996,22(2)：214-219
8.胡琼,李云昌,梅德圣,等.属间体细胞杂交创建甘蓝型油菜细胞质雄性不育系及其鉴定.中国农业科学,2004,37(3)：333-338
9.桓明辉,陈飞,吴红艳,等.不吸水链霉菌基因文库的构建及初步筛选.生物技术通报,2009,1：134-137
10.黄进勇,胡琼,魏文辉,等.甘蓝型油菜新型恢复系Nsa恢1及其后代的GISH分析.武汉大学学报,2008,54(4)：467-471
11.黄永菊,李云昌,陈军,等.新疆野生油菜芥酸含量的遗传分析.中国油料作物学报,1999,21(4)：15-16,20
12.蒋三亮,张思仲.用地高辛配基标记DNA探针进行菌落原位杂交.华西医大学报,1992,23(5)：245-247
13.焦清亮,王兆木.新疆野生油菜不育材料的发现与研究初报.新疆农业科学,2001,38(1)：39-40
14.李建平,张燕红,王冬梅,等.一种改良的Southern印迹杂交方法.新疆农业科学,2007,44(3)：116-118
15.李谨,杨公社,陈创夫,等.新疆细毛羊2倍基因组BAC文库PCR筛选系统的构建与使用.分子细胞生物学报,2006,36(9)：489-494
16.梅德圣,李云昌,胡琼.甘蓝型油菜属间体细胞杂种雄性不育材料的研究.中国油料作物学报,2003,25(1)：72-75
17.孙盼盼.油菜恢复基因的分子标记及恢复系菌核病的抗性鉴定.[硕士学位论文].北京：中国农业科学院,2008
18.陶小平,李磊,李强,等.优化茵落原位杂交体系筛选野生稻基因组文库.生命科学研究,2008,12(4)：328-332
19.王道杰,王灏,扬翠玲,等.油菜雄性不育分子机理研究进展.中国农学通报,2001,17：43-46
20.王兆木,焦清亮,李艺.新疆野生油菜的考察和研究.新疆农业科学,1990,5：204-206
21.王国槐,官春云,陈社员.新疆野生油菜与野芥品质性状的比较研究.作物品种资源,1999,1：37-38
22.王永飞,马三梅,刘翠平,等.分子标记在植物遗传育种中的应用原理及现状.西北农林科技大学学报,29：106-113
23.王振东,孙仓,王惠.不同方法从大豆不同组织中提取基因组DNA效果的比较.大豆科学,2008,27(1)：42-46
24.吴新东,陈芳,李鑫,等.中国美利奴细毛羊BAC文库的三维PCR筛选.生物工程学报,2008,24(10)：1828-1831
25.肖成汉.新疆野生油菜和西德黑芥核型的比较研究.华中农业大学学报,1989,8(4)：297-301
26.肖成汉,肖红.新疆野生油菜、黑芥、芥菜型和白菜型油菜酸性磷酸化酶同工酶的比较研究.中国油料,1993,3：12-14
27.尹经章.油菜杂种优势利用的现状与展望(综述).新疆农业大学学报,2002,25：40-46
28.战爱斌,胡景杰,胡晓丽,等.富集文库-菌落原位杂交法筛选栉孔扇贝的微卫星标记.水产学报,2008,32(3)：353-361
29.周长发,张锐,张晓,等.地高辛随机引物法标记探针Southern杂交技术优化.中国农业科技导报,2009,11(4)：123-128
30.Aert R, Sagi L, Volckaert G. Gene content and density in banana (Musa acuminata) as revealed by genomic sequencing of BAC clones.Theor Appl Genet,2004, 109:129-139
31.Arumuganathan K, Earle E D. Nuclear DNA content of some important plant species. Plant Molecular Biology Reporter,1991,9:208-218
32.Ahn S,Tanksley S D. Comparative linkage maps of the rice and maize genomes. Proc Natl Acad Sci USA.1993.90:7980-7984
33.Ausubel F M, Brent R, Kingston R E, et.al.《Short Protocols in Molecular Biology》, 3rd edition, John Wiley & Sons, Inc,1995
34. Bentolila S,Alfonso A A, Hanson M R. A pentatricopeptide repeat-containing gene restores fertility to cytoplasmic male-sterile plants. Proc Natl Acad Sci USA,2002, 99:10887-10892
35.Birren B.Cloning Systems In Genome Analysis:A laboratory manual. Cold Spring Harbor. New York:Cold Spring Harbor Laboratory Press,1997.242-295
36.Budar F, Touzet P, Paepe R D.The nucleo-mitochondrial conflict in cytoplasmic male sterilities revisited. Genetica,2003,117:3-16
37.Budiman M A, Mao L, Wood T C,et al.A Deep-Coverage Tomato BAC Library and Prospects Toward Development of an STC Framework for Genome Sequencing. Genome Res.2000,10:129-136
38.Burke D T,Carle G F,Olson M.Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors.Science,1987,236:806-812
39.Chumakov I M, Rigault P, Le Gall I, et al.A YAC contig map of the human genome. Nature,1995,377(6547):175-297
40.Collins J, Hohn B.Cosmids:a type of plasmid gene cloning vector that is packageable in vitro in bacterio phage lambda heads. Proc Natl Acad Sci USA,1978,75(9): 4242-4246
41 Cui X Q, Wise R P,Schnable P S.The rf2 nuclear restorer gene of male-sterile T-cytoplasm maize. Science,1996,272:1334-1336
42.Desloire S,Gherbi H, Laloui W, et al. Identification of the fertility restoration locus, Rf0,in radish, as a member of the pentatricopeptide-repeat protein family. EMBO Rep,2003,4:588-594
43.Frary A, Hamillon C M.Efficiency and stability of high molecular weight DNA transformation:an analysis in tomato.Transgenic Research,2001,10:121-132
44.Friiters A C, Wang Z, Van Damine M, et al.Construction of a BAC library containing large EcoR Ⅰ and Hind Ⅲ genomic fragments of lettuce. Theor Appl Genet,1997,94:390-399
45.Fu T D, Yang G S,Tu J X, et al. The present and future of rapeseed production in China. Chin Oil Fat,2003,28:11-13
46.Green E D, Riethman H C,Dutchik J E, et al.Detection and characterization of chimeric yeast artificial-chromosome clones.Genomics,1991,11:658-669
47. Goss S J, Harris H. New method for mapping genes in human chromosomes. Nature, 1975,255:680-684
48.Gregory S G, Sekhon M, Schein J, et al. A physical map of the mouse genome. Nature, 2002,418:743-750
49.Hamillon C M. A binary-BAC system for plant transformation with high-molecular-weight DNA. Gene,1997,200:107-116
50. Harrington J J, Van Bokkelen G, Mays R W, et al. Formation of de novo centromeres and construction of first-generation human artificial microchromosomes.Nat Genet, 1997,15:345-355
51.Harushima Y, Yano M, Shomura A, et al.A high-density rice genetic linkage map with 2275 markers using a single F2 population. Genetics,1998,148:479-494
52.HE Cong Fen, Takao Komatsuda. PCR-based Screening BAC Library and Direct End Sequencing of BAC Clones. Acta Genetica Sinic.2004,31(11):1262-1267
53.Henning K A, Novotny E A, Compton S T, et al.Human artificial chromosomes generated by modification of a yeast artificial chromosome containing both human alpha satellite and single-copy DNA sequences. Proc Natl Acad Sci USA,1999,96: 592-597
54.Hess M, Goldammer T, Gelhaus A et al. Physical assignment of the bovine MHC class Iia and class Ⅱb genes.Cytogenet Cell Genet,1999,85:244-247
55.Hosoda F, Nishimura S,Uchida H, et al. An F factor based cloning system for large DNA fragments. Nucleic Acids Research,1990,18:3863-3869
56.Hu Q, Andersen S B, Dixelias C, et al.Production of Fertile Intergeneric Somatic Hybrids between Brassica napus and Sinapis arvensis for the Enrichment of the Rapeseed Gene Pool.Plant Cell Rep,2002,21(2):147-152
57.Ratnayaka I, Baga M, Fowler D B,et al. Construction and Characterization of a BAC Library of a Cold-Tolerant Hexaploid Wheat Cultivar. Crop Science,2005,45: 1571-1577
58.Ioannou P A, Amemiya C T, Games J, et al.A new bacteriophage P1 derived vector for the propagation of large human DNA fragments. Nature Genet,1994,6:84-89
59.Iwabuchi M, Koizuka N, Fujimoto H, et al. Identification and expression of the kosena radish (Raphanus sativus cv. Kosena) homol Ogue of the Ogura radish CMS-associated gene, orf138. Plant Mol Biol,1999,39:183-188
60.Joobeur T, Gusmini G, Zhang X, et al. Construction of a watermelon BAC library and identification of SSRs anchored to melon or Arabidopsis genomes. Theor Appl Genet,2006,112:1553-1562
61.Kim U J, Birren B,Slepak T, et al.Construction and characterization of a human bacterial artificial chromosome library. Genomics,1996,34:213-218
62.Koizuka N, Imai R, Iwabuchi M, et al.Genetic analysis of fertility restoration and accumulation of ORF125 mitochondrial protein in the kosena radish (Raphanus sativus cv. Kosena) and a Brassica napus restorer line. Theor Appl Genet,2000,100: 949-955
63.Koizuka N, Imai R, Fujimoto H, et al.Genetic characterization of a pentatricopeptide repeat protein gene, orf687 that restores fertility in the cytoplasmic male-sterile Kosena radish. Plant J,2003,34:407-415
64.Komori T, Ohta S,Murai N. et al.Map-based cloning of a fertility restorer gene,Rf-1, in rice(Oryza sativa L.).Plant J,2004,37:315-325
65.Kurata N, Umehara Y,Tanoue H, et al.Physical mapping of the rice genome with YAC clones.Plant Mol Biol,1997,35:101-113
66.Leroy T, Marraccini P,Dufour M, et al.Construction and characterization of a Coffea canephora BAC library to study the organization of sucrose biosynthesis genes. Theor Appl Genet,2005,111:1032-1041
67.L'Homme Y, Brown G G. Organization differences between cytoplasmic male sterile and fertile Brassica mitochondrial genomes are confined to a single transposed locus. Nucleic Acids Res,1993,21:1903-1909
68.L'Homme Y, Stahl R, Li X Q, et al.Brassica nap cytoplasmic male sterility is associated with expression of an mtDNA region containing a chimeric gene similar to the Pol CMS-associated orf224 gene. Curr Genet,1997,31:325-335
69.Lijavetzky D, Muzzi G, Wicker T, et al.Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat. Genome, 1999,42:1176-1182
70.Liu Y G, Fukaki H, Yanai Y, et al. Complementation of plant mutants withlarge genomic DNA fragments by a transformation-competentartificial chromosome vector accelerates positional cloning. Proc Natl Acad Sci. USA,1999,96,6535-6540
71.Luo M Z, Wing R. An improved method for plant BAC library construction. Methods in Molecular Biology,2003,236:3-20
72.Mozo T, Fischer S,Shizuya H, et al.Construction and characterization of the IGF Arabidopsis BAC library. Mol Gen Genet,1998,258:562-570
73.Mozo T, Dewar K, Dunn P, et al.A complete BAC-based physical map of the Arabidopsis thaliana genome.Nature Genet,1999,22:271-275
74.Murray N E, Murray K. Manipulation of restriction targets in phage lambda to form receptor chromosomes for DNA fragments. Nature,1974,251(5475):476-481
75.Murry A W, Szostak J W. Construction of artificial chromosome in yeast. Nature, 1983,305:189-193
76.Nam Y W, Lee J R, Song K H, et al.Construction of two BAC librariesfrom cucumber(Cucumis sativus L.) and identification of clones linked to yield component quantitative trait loci.Theor Appl Genet,2005,111:150-161
77.Neil D L, Villasante A, Fisher RB, et al Structural instability of human tandemly repeated DNA sequences cloned in yeast artificial chromosome vectors.Nucl Acids Res,1990,18:1421-1428
78.Osoegawa K.,Woon P Y, Zhao B, et al.An improved approach for construction of bacterial artificial chromosome libraries.Genomics,1998,52:1-8
79.Peterson D G, Tomkins J P,Frisch D A, et al.Construction of plant bacterial artificial chromosome (BAC)libraries:An illustrated guide. Journal of Agricultural Genomics, 2000,5:1-10
80. Sambrook J, Fritsch E F, Maniatis T. Molecular cloning, a laboratory manual. Cold Spring Harbor, Cold Spring Harbor Laboratory Press,1989
81.Schlessinger D.Yeast artificial chromosomes:tools for mapping and analysis of complex genomes. Trends Genet,1990,6:248-258
82.Shizuya H, Birren B,Kim U J, et.al.Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector. Proceedings of the National Academy of Sciences of the United States of America, 1992,89:8794-8797
83.Asakawa S,Abe I, Kudoh Y, et al.Human BAC library:construction and rapid screening.Gene,1997,191:69-79
84.Shultz J L, Kazi S,Bashir R, et al.The development of BAC-end sequence-based microsatellite markers and placement in the physical and genetic maps of soybean, Theor Appl Genet,2007,114:1081-1090
85.Smajs D, McKevitt M, Wang L, et al. BAC Library of T. pallidum DNA in E. Coli. Genome Res,2002,12:515-522
86.Sternberg N. Bacteriophage Pl cloning system for the isolation amptification and recovery of DNA fragments as large as 100 kilobase pairs. Proc Natl Acad Sci USA, 1990,87:103-107
87. Stiewe G, Witt U, Hansen S,et al. Natural and experimental evolution of CMS for rapeseed breeding. Adv Plant Breeding,1995,18:59-76
88.Strong S J, Ohtal Y, Litman G W, et al. Marked improvement of PAC and BAC cloning isachieved using electroelution of pulsed-field gel-separated partial digests of genomic DNA. Nucleic Acids Research,1997,19 (25):3959-3961
89.Woo S S,Rastogi V K.,Zhang H B,et al. Isolation of Megabase-Size DNA from Sorghum and Applications for Physical Mapping and Bacterial and Yeast Artificial Chromosome Library Construction. Plant Molecular Biology Reporter.1995,13(1): 82-94
90.Telzur N, Abbo S,Myslabodski D, et al.Modified CTAB Procedure for DNA Isolation from Epiphytic Cacti of the Genera Hylocereus and Selenicereus (Cactaceae). Plant Molecular Biology Reporter,1999,17:249-254
91.Tomkins J P, Mahalingam R, Smith H,et al. A bacterial artificial chromosome library for soybean PI 437654 and identification of clones associated with cyst nematode resistance.Plant Molecular Biology,1999,41:25-32
92.Wang G L, Holsten T E, Song W Y, et al. Construction of a rice bacterial artificial chromosome library and identification ofclones linked to the Xa-21 disease resistance locus.Plant J.1995,7:525-533
93.Wang Y F,Ma S M, Wang M, et al. Sequence analysis of the gene correlated with cytoplasmic male sterility in rapeseed(Brassica napus L.)Polima and Shaan 2A. Chin Sci Bull,2002,47:122-126
94.Woo S S,Rastogi V K, Zhang H B,et al. Isolation of Megabase-Size DNA from Sorghum and Applications for Physical Mapping and Bacterial and Yeast Artificial Chromosome Library Construction. Plant Molecular Biology Reporter,1995,13(1): 82-94
95.Xu H L, Qian Y P, Nie W H,et al. Construction, characterization and chromosomal mapping of bacterial artificial chromosome (BAC) library of Yunnan snub-nosed monkey (Rhinopithecus bieti).Chromosome Research,2004,12:251-262
96.Zhang H B,Martin G B,Tanksley S D, et al.Map-based cloning in crop plants: tomato as a model system U. Isolation and characterization of a set of overlapping yeast artificial chromosomes encompassing the jointless locus. Mol Gen Genet,1994, 244:613-621
97.Zhang H B,Zhao X, Ding X, et al.Preparation of megabase-size DNA from plant nuclei.Plant J,2003,7:175-184
98.Zhang H B,Choi S,Woo S S,et al. Construction and characterization of two rice bacterial artificial chromosome libraries from the parents of a permanent recombinant inbred mapping population. Molecular Breeding,1996,2:11-24
2.陈庆祥.一种2n=18的芥菜型野生油菜.新疆农业科学,1986,1：18-19
3.陈书元,晁金泉,徐有明,等.甘蓝型油菜—新疆野生油菜二体异附加系BAC文库的构建.中国油料作物学报,2008,30(2)：137-142
4.程计华,李云昌,胡琼,等.油菜野芥NSa细胞质雄性不育系的特异性分子鉴定.作物学报,2008,34(11)：1946-1952
5.杜伟,王兆木,孙卫琳,等.新疆野生油菜体细胞染色体核型的研究.西北农业学报,1992,1(2)：8-10
6.杜伟,王兆木,孙卫琳,等.新疆野生油菜体细胞染色体数观察及统计.新疆农业科学,1992,4：156-157
7.官春云.新疆野生油菜与野芥(Sinapis arvensis L.)遗传性状的比较研究.作物学报,1996,22(2)：214-219
8.胡琼,李云昌,梅德圣,等.属间体细胞杂交创建甘蓝型油菜细胞质雄性不育系及其鉴定.中国农业科学,2004,37(3)：333-338
9.桓明辉,陈飞,吴红艳,等.不吸水链霉菌基因文库的构建及初步筛选.生物技术通报,2009,1：134-137
10.黄进勇,胡琼,魏文辉,等.甘蓝型油菜新型恢复系Nsa恢1及其后代的GISH分析.武汉大学学报,2008,54(4)：467-471
11.黄永菊,李云昌,陈军,等.新疆野生油菜芥酸含量的遗传分析.中国油料作物学报,1999,21(4)：15-16,20
12.蒋三亮,张思仲.用地高辛配基标记DNA探针进行菌落原位杂交.华西医大学报,1992,23(5)：245-247
13.焦清亮,王兆木.新疆野生油菜不育材料的发现与研究初报.新疆农业科学,2001,38(1)：39-40
14.李建平,张燕红,王冬梅,等.一种改良的Southern印迹杂交方法.新疆农业科学,2007,44(3)：116-118
15.李谨,杨公社,陈创夫,等.新疆细毛羊2倍基因组BAC文库PCR筛选系统的构建与使用.分子细胞生物学报,2006,36(9)：489-494
16.梅德圣,李云昌,胡琼.甘蓝型油菜属间体细胞杂种雄性不育材料的研究.中国油料作物学报,2003,25(1)：72-75
17.孙盼盼.油菜恢复基因的分子标记及恢复系菌核病的抗性鉴定.[硕士学位论文].北京：中国农业科学院,2008
18.陶小平,李磊,李强,等.优化茵落原位杂交体系筛选野生稻基因组文库.生命科学研究,2008,12(4)：328-332
19.王道杰,王灏,扬翠玲,等.油菜雄性不育分子机理研究进展.中国农学通报,2001,17：43-46
20.王兆木,焦清亮,李艺.新疆野生油菜的考察和研究.新疆农业科学,1990,5：204-206
21.王国槐,官春云,陈社员.新疆野生油菜与野芥品质性状的比较研究.作物品种资源,1999,1：37-38
22.王永飞,马三梅,刘翠平,等.分子标记在植物遗传育种中的应用原理及现状.西北农林科技大学学报,29：106-113
23.王振东,孙仓,王惠.不同方法从大豆不同组织中提取基因组DNA效果的比较.大豆科学,2008,27(1)：42-46
24.吴新东,陈芳,李鑫,等.中国美利奴细毛羊BAC文库的三维PCR筛选.生物工程学报,2008,24(10)：1828-1831
25.肖成汉.新疆野生油菜和西德黑芥核型的比较研究.华中农业大学学报,1989,8(4)：297-301
26.肖成汉,肖红.新疆野生油菜、黑芥、芥菜型和白菜型油菜酸性磷酸化酶同工酶的比较研究.中国油料,1993,3：12-14
27.尹经章.油菜杂种优势利用的现状与展望(综述).新疆农业大学学报,2002,25：40-46
28.战爱斌,胡景杰,胡晓丽,等.富集文库-菌落原位杂交法筛选栉孔扇贝的微卫星标记.水产学报,2008,32(3)：353-361
29.周长发,张锐,张晓,等.地高辛随机引物法标记探针Southern杂交技术优化.中国农业科技导报,2009,11(4)：123-128
30.Aert R, Sagi L, Volckaert G. Gene content and density in banana (Musa acuminata) as revealed by genomic sequencing of BAC clones.Theor Appl Genet,2004, 109:129-139
31.Arumuganathan K, Earle E D. Nuclear DNA content of some important plant species. Plant Molecular Biology Reporter,1991,9:208-218
32.Ahn S,Tanksley S D. Comparative linkage maps of the rice and maize genomes. Proc Natl Acad Sci USA.1993.90:7980-7984
33.Ausubel F M, Brent R, Kingston R E, et.al.《Short Protocols in Molecular Biology》, 3rd edition, John Wiley & Sons, Inc,1995
34. Bentolila S,Alfonso A A, Hanson M R. A pentatricopeptide repeat-containing gene restores fertility to cytoplasmic male-sterile plants. Proc Natl Acad Sci USA,2002, 99:10887-10892
35.Birren B.Cloning Systems In Genome Analysis:A laboratory manual. Cold Spring Harbor. New York:Cold Spring Harbor Laboratory Press,1997.242-295
36.Budar F, Touzet P, Paepe R D.The nucleo-mitochondrial conflict in cytoplasmic male sterilities revisited. Genetica,2003,117:3-16
37.Budiman M A, Mao L, Wood T C,et al.A Deep-Coverage Tomato BAC Library and Prospects Toward Development of an STC Framework for Genome Sequencing. Genome Res.2000,10:129-136
38.Burke D T,Carle G F,Olson M.Cloning of large segments of exogenous DNA into yeast by means of artificial chromosome vectors.Science,1987,236:806-812
39.Chumakov I M, Rigault P, Le Gall I, et al.A YAC contig map of the human genome. Nature,1995,377(6547):175-297
40.Collins J, Hohn B.Cosmids:a type of plasmid gene cloning vector that is packageable in vitro in bacterio phage lambda heads. Proc Natl Acad Sci USA,1978,75(9): 4242-4246
41 Cui X Q, Wise R P,Schnable P S.The rf2 nuclear restorer gene of male-sterile T-cytoplasm maize. Science,1996,272:1334-1336
42.Desloire S,Gherbi H, Laloui W, et al. Identification of the fertility restoration locus, Rf0,in radish, as a member of the pentatricopeptide-repeat protein family. EMBO Rep,2003,4:588-594
43.Frary A, Hamillon C M.Efficiency and stability of high molecular weight DNA transformation:an analysis in tomato.Transgenic Research,2001,10:121-132
44.Friiters A C, Wang Z, Van Damine M, et al.Construction of a BAC library containing large EcoR Ⅰ and Hind Ⅲ genomic fragments of lettuce. Theor Appl Genet,1997,94:390-399
45.Fu T D, Yang G S,Tu J X, et al. The present and future of rapeseed production in China. Chin Oil Fat,2003,28:11-13
46.Green E D, Riethman H C,Dutchik J E, et al.Detection and characterization of chimeric yeast artificial-chromosome clones.Genomics,1991,11:658-669
47. Goss S J, Harris H. New method for mapping genes in human chromosomes. Nature, 1975,255:680-684
48.Gregory S G, Sekhon M, Schein J, et al. A physical map of the mouse genome. Nature, 2002,418:743-750
49.Hamillon C M. A binary-BAC system for plant transformation with high-molecular-weight DNA. Gene,1997,200:107-116
50. Harrington J J, Van Bokkelen G, Mays R W, et al. Formation of de novo centromeres and construction of first-generation human artificial microchromosomes.Nat Genet, 1997,15:345-355
51.Harushima Y, Yano M, Shomura A, et al.A high-density rice genetic linkage map with 2275 markers using a single F2 population. Genetics,1998,148:479-494
52.HE Cong Fen, Takao Komatsuda. PCR-based Screening BAC Library and Direct End Sequencing of BAC Clones. Acta Genetica Sinic.2004,31(11):1262-1267
53.Henning K A, Novotny E A, Compton S T, et al.Human artificial chromosomes generated by modification of a yeast artificial chromosome containing both human alpha satellite and single-copy DNA sequences. Proc Natl Acad Sci USA,1999,96: 592-597
54.Hess M, Goldammer T, Gelhaus A et al. Physical assignment of the bovine MHC class Iia and class Ⅱb genes.Cytogenet Cell Genet,1999,85:244-247
55.Hosoda F, Nishimura S,Uchida H, et al. An F factor based cloning system for large DNA fragments. Nucleic Acids Research,1990,18:3863-3869
56.Hu Q, Andersen S B, Dixelias C, et al.Production of Fertile Intergeneric Somatic Hybrids between Brassica napus and Sinapis arvensis for the Enrichment of the Rapeseed Gene Pool.Plant Cell Rep,2002,21(2):147-152
57.Ratnayaka I, Baga M, Fowler D B,et al. Construction and Characterization of a BAC Library of a Cold-Tolerant Hexaploid Wheat Cultivar. Crop Science,2005,45: 1571-1577
58.Ioannou P A, Amemiya C T, Games J, et al.A new bacteriophage P1 derived vector for the propagation of large human DNA fragments. Nature Genet,1994,6:84-89
59.Iwabuchi M, Koizuka N, Fujimoto H, et al. Identification and expression of the kosena radish (Raphanus sativus cv. Kosena) homol Ogue of the Ogura radish CMS-associated gene, orf138. Plant Mol Biol,1999,39:183-188
60.Joobeur T, Gusmini G, Zhang X, et al. Construction of a watermelon BAC library and identification of SSRs anchored to melon or Arabidopsis genomes. Theor Appl Genet,2006,112:1553-1562
61.Kim U J, Birren B,Slepak T, et al.Construction and characterization of a human bacterial artificial chromosome library. Genomics,1996,34:213-218
62.Koizuka N, Imai R, Iwabuchi M, et al.Genetic analysis of fertility restoration and accumulation of ORF125 mitochondrial protein in the kosena radish (Raphanus sativus cv. Kosena) and a Brassica napus restorer line. Theor Appl Genet,2000,100: 949-955
63.Koizuka N, Imai R, Fujimoto H, et al.Genetic characterization of a pentatricopeptide repeat protein gene, orf687 that restores fertility in the cytoplasmic male-sterile Kosena radish. Plant J,2003,34:407-415
64.Komori T, Ohta S,Murai N. et al.Map-based cloning of a fertility restorer gene,Rf-1, in rice(Oryza sativa L.).Plant J,2004,37:315-325
65.Kurata N, Umehara Y,Tanoue H, et al.Physical mapping of the rice genome with YAC clones.Plant Mol Biol,1997,35:101-113
66.Leroy T, Marraccini P,Dufour M, et al.Construction and characterization of a Coffea canephora BAC library to study the organization of sucrose biosynthesis genes. Theor Appl Genet,2005,111:1032-1041
67.L'Homme Y, Brown G G. Organization differences between cytoplasmic male sterile and fertile Brassica mitochondrial genomes are confined to a single transposed locus. Nucleic Acids Res,1993,21:1903-1909
68.L'Homme Y, Stahl R, Li X Q, et al.Brassica nap cytoplasmic male sterility is associated with expression of an mtDNA region containing a chimeric gene similar to the Pol CMS-associated orf224 gene. Curr Genet,1997,31:325-335
69.Lijavetzky D, Muzzi G, Wicker T, et al.Construction and characterization of a bacterial artificial chromosome (BAC) library for the A genome of wheat. Genome, 1999,42:1176-1182
70.Liu Y G, Fukaki H, Yanai Y, et al. Complementation of plant mutants withlarge genomic DNA fragments by a transformation-competentartificial chromosome vector accelerates positional cloning. Proc Natl Acad Sci. USA,1999,96,6535-6540
71.Luo M Z, Wing R. An improved method for plant BAC library construction. Methods in Molecular Biology,2003,236:3-20
72.Mozo T, Fischer S,Shizuya H, et al.Construction and characterization of the IGF Arabidopsis BAC library. Mol Gen Genet,1998,258:562-570
73.Mozo T, Dewar K, Dunn P, et al.A complete BAC-based physical map of the Arabidopsis thaliana genome.Nature Genet,1999,22:271-275
74.Murray N E, Murray K. Manipulation of restriction targets in phage lambda to form receptor chromosomes for DNA fragments. Nature,1974,251(5475):476-481
75.Murry A W, Szostak J W. Construction of artificial chromosome in yeast. Nature, 1983,305:189-193
76.Nam Y W, Lee J R, Song K H, et al.Construction of two BAC librariesfrom cucumber(Cucumis sativus L.) and identification of clones linked to yield component quantitative trait loci.Theor Appl Genet,2005,111:150-161
77.Neil D L, Villasante A, Fisher RB, et al Structural instability of human tandemly repeated DNA sequences cloned in yeast artificial chromosome vectors.Nucl Acids Res,1990,18:1421-1428
78.Osoegawa K.,Woon P Y, Zhao B, et al.An improved approach for construction of bacterial artificial chromosome libraries.Genomics,1998,52:1-8
79.Peterson D G, Tomkins J P,Frisch D A, et al.Construction of plant bacterial artificial chromosome (BAC)libraries:An illustrated guide. Journal of Agricultural Genomics, 2000,5:1-10
80. Sambrook J, Fritsch E F, Maniatis T. Molecular cloning, a laboratory manual. Cold Spring Harbor, Cold Spring Harbor Laboratory Press,1989
81.Schlessinger D.Yeast artificial chromosomes:tools for mapping and analysis of complex genomes. Trends Genet,1990,6:248-258
82.Shizuya H, Birren B,Kim U J, et.al.Cloning and stable maintenance of 300-kilobase-pair fragments of human DNA in Escherichia coli using an F-factor-based vector. Proceedings of the National Academy of Sciences of the United States of America, 1992,89:8794-8797
83.Asakawa S,Abe I, Kudoh Y, et al.Human BAC library:construction and rapid screening.Gene,1997,191:69-79
84.Shultz J L, Kazi S,Bashir R, et al.The development of BAC-end sequence-based microsatellite markers and placement in the physical and genetic maps of soybean, Theor Appl Genet,2007,114:1081-1090
85.Smajs D, McKevitt M, Wang L, et al. BAC Library of T. pallidum DNA in E. Coli. Genome Res,2002,12:515-522
86.Sternberg N. Bacteriophage Pl cloning system for the isolation amptification and recovery of DNA fragments as large as 100 kilobase pairs. Proc Natl Acad Sci USA, 1990,87:103-107
87. Stiewe G, Witt U, Hansen S,et al. Natural and experimental evolution of CMS for rapeseed breeding. Adv Plant Breeding,1995,18:59-76
88.Strong S J, Ohtal Y, Litman G W, et al. Marked improvement of PAC and BAC cloning isachieved using electroelution of pulsed-field gel-separated partial digests of genomic DNA. Nucleic Acids Research,1997,19 (25):3959-3961
89.Woo S S,Rastogi V K.,Zhang H B,et al. Isolation of Megabase-Size DNA from Sorghum and Applications for Physical Mapping and Bacterial and Yeast Artificial Chromosome Library Construction. Plant Molecular Biology Reporter.1995,13(1): 82-94
90.Telzur N, Abbo S,Myslabodski D, et al.Modified CTAB Procedure for DNA Isolation from Epiphytic Cacti of the Genera Hylocereus and Selenicereus (Cactaceae). Plant Molecular Biology Reporter,1999,17:249-254
91.Tomkins J P, Mahalingam R, Smith H,et al. A bacterial artificial chromosome library for soybean PI 437654 and identification of clones associated with cyst nematode resistance.Plant Molecular Biology,1999,41:25-32
92.Wang G L, Holsten T E, Song W Y, et al. Construction of a rice bacterial artificial chromosome library and identification ofclones linked to the Xa-21 disease resistance locus.Plant J.1995,7:525-533
93.Wang Y F,Ma S M, Wang M, et al. Sequence analysis of the gene correlated with cytoplasmic male sterility in rapeseed(Brassica napus L.)Polima and Shaan 2A. Chin Sci Bull,2002,47:122-126
94.Woo S S,Rastogi V K, Zhang H B,et al. Isolation of Megabase-Size DNA from Sorghum and Applications for Physical Mapping and Bacterial and Yeast Artificial Chromosome Library Construction. Plant Molecular Biology Reporter,1995,13(1): 82-94
95.Xu H L, Qian Y P, Nie W H,et al. Construction, characterization and chromosomal mapping of bacterial artificial chromosome (BAC) library of Yunnan snub-nosed monkey (Rhinopithecus bieti).Chromosome Research,2004,12:251-262
96.Zhang H B,Martin G B,Tanksley S D, et al.Map-based cloning in crop plants: tomato as a model system U. Isolation and characterization of a set of overlapping yeast artificial chromosomes encompassing the jointless locus. Mol Gen Genet,1994, 244:613-621
97.Zhang H B,Zhao X, Ding X, et al.Preparation of megabase-size DNA from plant nuclei.Plant J,2003,7:175-184
98.Zhang H B,Choi S,Woo S S,et al. Construction and characterization of two rice bacterial artificial chromosome libraries from the parents of a permanent recombinant inbred mapping population. Molecular Breeding,1996,2:11-24