摘要
目的:构建光滑假丝酵母ATCC2001菌株PDR1基因敲除突变株。方法:用PRODIGE同源重组技术,以pY24GAL10为模板,设计80 bp长引物合成PDR1-URA3基因敲除组件,并将其转化入URA3已发生突变的ATCC2001/ura3菌株,经SD-URA选择性平板筛选后获得稳定敲除PDR1的ATCC2001/ura3 pdr1Δ菌株。结果:成功构建光滑假丝酵母PDR1基因敲除菌株ATCC2001/ura3 pdr1Δ。结论:此法可用于精确敲除光滑假丝酵母目标基因。ATCC2001/ura3 pdr1Δ菌株的构建将为进一步研究该基因在光滑假丝酵母中的功能及其作用机制奠定基础。
Objective： To construct a PDR1 knock - out strain of Candida glabrata ATCC2001. Method： Disrupting PDR1 gene by means of PRODIGE homologous recombination. Gene knock - out components PDR1 - URA3 was amplified with 80 bp long primers and pY24GAL10 as template, and transformed it into ATCC2001/ura3 strain which ura3 mutation has occurred. Then, ATCC2001/ura3 pdrl A strain was constructed by screening of SD- URA selectivity plate. Result：A PDR1 knock- out strain of ATCC2001/ura3 pdrl Awas suc- cessfully constructed. Conclusion：The gene knock - out method is stable with a good efficiency, and ATCC2001/ura3 pdrl A strain can be used for detailed gene function studies of PDR1 in Candida glabrata.
出处
《生物技术》
CAS
CSCD
北大核心
2013年第4期43-46,共4页
Biotechnology
