| 引用本文： |
-
陆雁,王青艳,朱绮霞,秦艳,申乃坤,廖思明,谢能中,黄日波.枯草芽孢杆菌高效转化及其转化子验证方法[J].广西科学院学报,2012,28(2):117-119. [点击复制]
- LU Yan,WANG Qing-yan,ZHU Qi-xia,QIN Yan,SHEN Nai-kun,LIAO Si-ming,XIE Neng-zhong,HUANG Ri-bo.Method for Enhancing the Transformation Efficiency in Bacillus subtilis[J].Journal of Guangxi Academy of Sciences,2012,28(2):117-119. [点击复制]
|
|
| |
|
|
| 本文已被：浏览 413次 下载 1173次 |
码上扫一扫！ |
| 枯草芽孢杆菌高效转化及其转化子验证方法 |
|
陆雁1, 王青艳2, 朱绮霞2, 秦艳2, 申乃坤2, 廖思明1,2, 谢能中2, 黄日波1,2
|
|
|
| (1.广西大学生命科学与技术学院, 广西南宁 530004;2.广西科学院国家非粮生物质能源工程技术研究中心, 广西南宁 530007) |
|
| 摘要: |
| 用构建好的重组质粒转化枯草芽孢杆菌(Bacillus subtilis)WB600感受态细胞,进行不同时间预培养后,涂布于抗生素平板培养,研究一种更高效的转化方法。用载体与外源片段的连接产物转化WB600感受态细胞,进行最佳预培养时间培养后,涂布于抗生素平板培养,培养得到的转化子用PCR快速验证和双酶切鉴定,研究一种转化子的快速验证方法。 |
| 关键词: 枯草芽孢杆菌 转化方法 转化率 感受态细胞 |
| DOI： |
| 投稿时间：2012-04-25修订日期：2012-05-10 |
| 基金项目:广西自然科学基金重点项目(2010GXNSFD013030)、广西培养新世纪学术和技术带头人专项资金项目、广西科学研究与技术开发课题项目(桂科攻11107008-4)资助。 |
|
| Method for Enhancing the Transformation Efficiency in Bacillus subtilis |
|
LU Yan1, WANG Qing-yan2, ZHU Qi-xia2, QIN Yan2, SHEN Nai-kun2, LIAO Si-ming1,2, XIE Neng-zhong2, HUANG Ri-bo1,2
|
| (1.College of Life Science and Technology, Guangxi University, Nanning, Guangxi, 530004, China;2.National Engineering Research Center for Non-food Biorefinery, Guangxi Academy of Sciences, Nanning, Guangxi, 530007, China) |
| Abstract: |
| In order to study an effective method for transformation,Bacillus subtilis WB600 competent cells, transformed with pre-built recombinant plasmids, were precultured with different period of time and smeared to the plates containing antibiotics. In order to construct a fast and effective method for transformants verification, WB600 competent cells were transformed with vector and foreign DNA, preincubated with the optimal time, and smeared to the plates containing antibiotics. The transformants were verified with PCR and double restriction enzyme digestion. |
| Key words: Bacillus subtilis transformation approach transformation efficiency competent cell |
|
|
|
|
|
