CRISPR-Cas9系统与mazF介导的大片段删减法在酿酒酵母染色体大片段删减中的比较

Comparison of CRISRP-Cas9 system and mazF-mediated method for large deletions in Saccharomyces cerevisiae

摘要:

[目的]比较CRISPR-Cas9系统与mazF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%；mazF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；mazF法相对稳定，对目的基因无PAM位点要求。

关键词:酿酒酵母;大片段删减;CRISPR-Cas9;mazF;方法比较

Abstract:

[Objective] The objective of this research was to compare CRISRP-Cas9 system and mazF-mediated method for large deletions in Saccharomyces cerevisiae.[Methods] We made a 26.5 kb deletion from YKL072W to YKL061W 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 mazF-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 mazF-mediated method is stable and does not need the PAM sequence.

Key words:Saccharomyces cerevisiae;large deletion;CRISPR-Cas9;mazF;comparison