摘要
【目的】比较CRISPR-Cas9系统与maz F法这两种酿酒酵母染色体大片段删减方法。【方法】分别用上述两种方法删减了酿酒酵母长度为26.5 kb的染色体大片段YKL072W-YKL061W,并比较了两种方法的转化效率、敲除成功率。【结果】利用CRISPR-Cas9系统平均得到5个转化子,但正确率为100%;maz F法得到约100个转化子,正确率略低于前者,为93%。【结论】两种方法均能高效删减酿酒酵母染色体大片段,CRISPR-Cas9系统正确率较高,操作简便省时;maz F法相对稳定,对目的基因无PAM位点要求。
[Objective] The objective of this research was to compare CRISRP-Cas9 system and maz F-mediated method for large deletions in Saccharomyces cerevisiae. [Methods] We made a 26.5 kb deletion from YKL072 W to YKL061 W using the foresaid two methods. The two methods were analyzed from the perspective of transformation rate and accuracy of deletion. [Results] There were five colonies appeared on the plate in average using CRISRP-Cas9 system and all of these were correct. And 100 colonies were observed using the maz F-mediated method and the accuracy was 93%, a little bit lower than CRISPR-Cas9 system. [Conclusion] Both methods are good to make large deletion in Saccharomyces cerevisiae. The CRISRP-Cas9 system has a high accuracy and easy to use, the maz F-mediated method is stable and does not need the PAM sequence.
作者
吴玉珍
徐海津
白艳玲
张秀明
乔明强
Yuzhen Wu Haijin Xu Yanling Bai Xiuming Zhang Mingqiang Qiao(College of Life Sciences, Nankai University, Tianjin 300071, China Key Laboratory of Molecular Microbiology and Technology, Ministry of Education, Nankai University, Tianjin 300071, China)
出处
《微生物学报》
CAS
CSCD
北大核心
2017年第11期1604-1611,共8页
Acta Microbiologica Sinica
基金
国家自然科学基金(31470004)
国家"973计划"(2011CBA00802)~~
