| 同源重组法构建枯草芽孢杆菌dhbC基因缺失突变株和回复株 |
| |
| 引用本文： | 余贤美,林超,杨芩,李锐,贺春萍,郑服丛.同源重组法构建枯草芽孢杆菌dhbC基因缺失突变株和回复株[J].热带作物学报,2009,30(10):1517-1521. |
| |
| 作者姓名： | 余贤美 林超 杨芩 李锐 贺春萍 郑服丛 |
| |
| 作者单位： | 1. 中国热带农业科学院环境与植物保护研究所,海南儋州,571737;农业部热带农林有害生物入侵监测与控制重点开放实验室,海南儋州,571737;海南省热带农业有害生物检测监控重点实验室,海南儋州,571737
2. 中国热带农业科学院环境与植物保护研究所,海南儋州,571737;海南大学环境与植物保护学院,海南儋州,571737 |
| |
| 基金项目： | 中央级公益性科研院所基本科研业务费专项资助项目(No.2008hzslJ013;2008hzslJ014).国家科技支撑计划(No. 2007BAD48B04).公益性行业(农业)科研专项 |
| |
| 摘 要： | 为了验证dhbC基因的功能,以质粒pEGFP-NI为模板,通过PCR扩增获得新霉素抗性基因(neo~r)DNA片段,构建重组质粒pMD18-neo,电击转化法将pMD18-neo导入枯草芽孢杆菌Bacillus subtilis CAS15感受态细胞中,使neo'基因片段与dhbC基因片段发生置换,获得dhbC基因缺失突变株.再通过电击转化将dhbC 基因全长编码序列导入dhbC基因缺失突变株CAS15 dhbC-del,获得dhbC基因回复株CAS15 dhbC-com.经CAS平板法检测表明,CAS15 dhbC-del不能产生橘黄色晕圈,而CAS15 dhbC-tom产生与CAS一致的橘黄色晕圈,证明了dhbC基因与嗜铁索的合成密切相关.
|
| 关 键 词： | 枯草芽孢杆菌 同源重组 缺失突变 基因回复 |
Construction of Bacillus subtilis CAS15 dhbC- and dhbC+ Mutants by Using the Homologous Recombination Method |
| |
| Yu Xianmei,Lin Chao,Yang Qin,Li Rui,He Chunping and Zheng Fucong.Construction of Bacillus subtilis CAS15 dhbC- and dhbC+ Mutants by Using the Homologous Recombination Method[J].Chinese Journal of Tropical Crops,2009,30(10):1517-1521. |
| |
| Authors: | Yu Xianmei Lin Chao Yang Qin Li Rui He Chunping and Zheng Fucong |
| |
| Abstract: | neor DNA fragments were obtained by PCR using plasmid EGFP-Nlas templates, and the recombinant plasmid pMD 18-neo was constructed. The pMD 18-neo was transformed into the competent cell of B. sub tills CAS15 via electroporation, and dhbC deletion mutant CAS15 dhbC-del was obtained by replacing the dhbC gene with neor DNA fragments. Then the dhbC complement mutant CAS15 dhbC-com was obtained by retransforming the dhbC gene into CAS15 dhbC-del via electroporation. Finally, CAS15 dhbC-del and CAS15 dhbC-com were inoculated on CAS agar plate. The result showed that CAS15 dhbC-com produced a significant orange halo as CAS 15, while CAS 15 dhbC-del does not, which indicated that the dhbC gene plays an important role in the siderophore production of CAS15. |
| |
| Keywords: | Bacillus subtilis homologous recombination gene knockout deletion mutagenesis gene complementation |
| 本文献已被 维普 万方数据 等数据库收录！ |
| 点击此处可从《热带作物学报》浏览原始摘要信息 |
| 点击此处可从《热带作物学报》下载免费的PDF全文 |
|
