CLC number: Q812
On-line Access: 2019-10-09
Received: 2019-05-24
Revision Accepted: 2019-06-20
Crosschecked: 2019-09-12
Cited: 0
Clicked: 3767
Zhang-Qing Song, Zhi-Jun Liao, Ye-Feng Hu, Zheng Ma, Andreas Bechthold, Xiao-Ping Yu. Development and optimization of an intergeneric conjugation system and analysis of promoter activity in Streptomyces rimosus M527[J]. Journal of Zhejiang University Science B,in press.Frontiers of Information Technology & Electronic Engineering,in press.https://doi.org/10.1631/jzus.B1900270 @article{title="Development and optimization of an intergeneric conjugation system and analysis of promoter activity in Streptomyces rimosus M527", %0 Journal Article TY - JOUR
龟裂链霉菌M527接合转移体系的建立和优化以及启动子的活性分析创新点:有效的遗传转化系统和合适的启动子是链霉菌基因表达和基因工程的必要前提.由于链霉菌的遗传背景复杂,接合转移实验参数对菌种具有高度特异性.因此,本研究建立并优化了一套适用于S. rimosus M527的属间接合转移体系,并在此基础上分析比较了四个启动子的表达活性.这将为进一步遗传修饰S. rimosus M527奠定基础. 方法:以S. rimosus M527为研究对象,通过对接合转移体系中的一些重要的实验参数(如接合转移培养 基、供受体比例、热激温度和混合培养时长等)进行了优化.在此基础上,构建以gusA为报告基因的质粒,通过直观显色反应以及GUS酶活的定量检测,分析比较了四个启动子的表达活性. 结论:建立了一种有效的适用于S. rimosus M527接合转移体系,优化后的接合效率最高达3.05×10−5.分析测试的四个启动子中,合成启动子SPL-21表现出最高表达活性,比常用强组成型启动子permE*活性高出2.2倍,合成启动子SPL-57与permE*的活性无显著差异,内源启动子potrB的活性比permE*降低了50%. 关键词组: Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article
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