论文部分内容阅读
目的构建羊布氏菌超氧化物歧化酶(Superoxide dismutase,SOD)基因重组酿酒酵母菌株,并进行鉴定。方法 PCR扩增羊布氏菌16M株SOD基因,与psos载体连接,构建重组表达质粒psos-SOD,转化酿酒酵母菌cdc25H,并进行PCR鉴定、表型验证、自激活及定位情况检验。结果重组真核表达质粒psos-SOD经双酶切和测序证明构建正确;重组酵母菌经PCR可扩增出525 bp的SOD基因条带,其表型正常,无自激活宿主菌的作用,表达蛋白定位正确。结论已成功构建了羊布氏菌SOD基因重组酿酒酵母菌株,为研究布氏菌致病的分子机制奠定了基础。
Objective To construct the recombinant Saccharomyces cerevisiae strain of Superoxide dismutase (SOD) gene of B.brevis. Methods The SOD gene of 16M strain was amplified by PCR and ligated with psos vector. The recombinant plasmid psos-SOD was constructed and transformed into Saccharomyces cerevisiae cdc25H. The PCR products were identified by PCR, phenotypic verification, self-activation and localization test. Results The recombinant eukaryotic expression plasmid psos-SOD was confirmed by double enzyme digestion and sequencing. The recombinant yeast strain amplified a 525 bp SOD gene band by PCR, which showed normal phenotype and no self-activating host bacteria expression Protein localization is correct. Conclusion The recombinant Saccharomyces cerevisiae strain was successfully constructed, which laid the foundation for the study on the molecular mechanism of brucellosis.