关键词: 载体构建, 酿酒酵母, 同义突变

Abstract:

In order to further understand the relationship between the expression of APA1 and hydrogen sulfide production in
Saccharomyces cerevisiae, synonymous mutations were introduced into APA1. The sequence of APA1 gene was cloned from
S288c by PCR, connected with EGFP gene, and inserted into pY16 vector. The vector was used for synonymous mutations,
and mutations of APA1-1/2/3/4 were obtained. According to the sequencing results, the mutations were correct. Synonymous
mutations of APA1-1/2/3/4 were successfully acquired and could be used for further studies. Therefore, one-step sitedirected
mutagenesis is an effective method. The fluorescence intensity of Saccharomyces cerevisiae YS59 transformed with
pY16TEF1-EGFP-APA1-1/2/3/4 with fluorescence microscope confirmed that APA1 and EGFP were co-expressed.

Key words: vector construction, Saccharomyces cerevisiae, synonymous mutation