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| 棉花线粒体基因组细菌人工染色体文库的构建 | |
| 引用本文： | 代培红,孟志刚,罗淑萍,张锐,程海刚,郭三堆.棉花线粒体基因组细菌人工染色体文库的构建[J].新疆农业大学学报,2007,30(4):57-62. |
| 作者姓名： | 代培红 孟志刚 罗淑萍 张锐 程海刚 郭三堆 |
| 作者单位： | 1. 中国农业科学院,生物技术研究所,北京,100081;新疆农业大学,农学院,乌鲁木齐,830052 2. 中国农业科学院,生物技术研究所,北京,100081 3. 新疆农业大学,农学院,乌鲁木齐,830052 |
| 基金项目： | 国家高技术研究发展计划(863计划) |
| 摘 要： | 以P30A细胞质雄性不育系黄化苗为材料,采用蔗糖密度差速离心和不连续蔗糖密度梯度超速离心提取棉花线粒体,低熔点琼脂糖包埋和原位消化裂解的方法制备高分子量的线粒体DNA(mtDNA),经BamHⅠ,HindⅢ酶切和PCR扩增进行纯度鉴定,并对其进行部分酶切,酶切片段与载体连接,电击转入大肠杆菌DH10B中,成功的构建了棉花线粒体基因组BAC文库。该文库共挑取3 840个单克隆,平均插入片段大小为15.5 kb,覆盖率超过70倍,研究结果表明该基因文库具有较高的质量。 |
| 关 键 词： | 棉花 细胞质雄性不育 线粒体基因组(mtDNA) 细菌人工染色体 文库 |
| 文章编号： | 1007-8614（2007）04-0057-06 |
| 修稿时间： | 2007-05-08 |
Construction of BAC Library of Cotton Cytoplasmic Male Sterility (mt DNA) |
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| DAI Pei-hong,MENG Zhi-gang,LUO Shu-ping,ZHANG Rui,CHENG Hai-gang,GUO San-dui.Construction of BAC Library of Cotton Cytoplasmic Male Sterility (mt DNA)[J].Journal of Xinjiang Agricultural University,2007,30(4):57-62. | |
| Authors: | DAI Pei-hong MENG Zhi-gang LUO Shu-ping ZHANG Rui CHENG Hai-gang GUO San-dui |
| Institution: | 1. Biotechnology Reasearch Institute of CAAS, Beijing, 100081; 2. College of Agronomy, Xin jiang Agricultural University, Urumqi, 830052 |
| Abstract: | A procedure to isolate mtDNA from cotton seedlings has been developed.The protocol allows cotton mitochondrion of high purity and yield to be isolated by using differential centrifugation,discontinuous sucrose gradient centrifugation of sucrose density.High molecular weight mtDNA was prepared by embedding low melt point agrose and instu digesting and lysis.Then the mtDNA of cytoplasmic male sterility material P30A was partly digested by restriction endonuclease,and was ligated with the BAC vector through T4 ligase.Electroporating into E.coli DH10B,the BAC library of cotton mtDNA was successfully constructed.The library consisted of about 3 840 clones with insert size 15.5 kb.It was calculated that the library contains approximate 70 times of the cotton mitochondrial genome.It was demonstrated this genome library has high quality. |
| Keywords: | cotton cytoplasmic male sterility mitochondrial genome Bacterial Artificial Chromosome(BAC) library |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |