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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 knockout mutant of Candida glabra ATCC2001 strain. Methods: PDR1-URA3 gene knockout kit was designed by using PRODIGE homologous recombination technique and pY24GAL10 as a template. The 80 bp long primer was designed and transformed into the ATCC2001 / ura3 strain in which URA3 had been mutated. After the SD-URA selective plate The strain ATCC2001 / ura3 pdr1Δ stably knocked out PDR1 was obtained after screening. Results: The Candida glabrata PDR1 knockout strain ATCC2001 / ura3 pdr1Δ was successfully constructed. Conclusion: This method can be used to accurately knock out the Candida glabrata target gene. The construction of ATCC2001 / ura3 pdr1Δ strain will lay the foundation for further study of the function and mechanism of this gene in Candida glabrata.