羊布氏菌SOD基因重组酿酒酵母菌株的构建及鉴定  

Construction and Identification of Recombinant Saccharomyces cerevisiae Strain for Superoxide Dismutase Gene of Brucella melitensis

作　　者：王景龙[1,2] 屈海龙[2] 杨延玲[1,2] 孙春辉[1,2] 王秀然[1,2] 郎需龙[2,3] 李晓艳[2,3] 卜昭阳[2,3] 王兴龙[2,3] 

机构地区：[1]吉林大学农学部畜牧兽医学院,长春130062 [2]军事医学科学院军事兽医研究所,长春130062 [3]吉林省人兽共患病控制与预防中心,长春130062

出　　处：《中国生物制品学杂志》2011年第6期639-642,共4页Chinese Journal of Biologicals

基　　金：国家自然科学基金(30972198/C180503)

摘　　要：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 and identify a recombinant Saccharomyces cerevisiae strain for superoxide dismutase（SOD） gene of Brucella melitensis.Methods The SOD gene of B.melitensis 16M strain was amplified by PCR and inserted into vector psos.The constructed recombinant plasmid was transformed to S.cerevisiae cdc25H,and the recombinants were identified by PCR,verified for phenotype,and tested for auto-activation and location.Results Both restriction analysis and sequencing proved that recombinant plasmid psos-SOD was constructed correctly.The SOD gene at a length of 525 bp was amplified by PCR from the recombinant S.cerevisiae which showed normal phenotype.The bait plasmid showed no auto-activation to host S.cerevisiae,and the expressed fusion protein was located correctly in the cytoplasma.Conclusion A recombinant S.cerevisiae strain for SOD gene of B.melitensis was successfully constructed,which laid a foundation of study on molecular mechanism of pathogenesis of brucella.

关 键 词：羊布氏菌 超氧化物歧化酶 酿酒酵母cdc25H 

分 类 号：R378.5[医药卫生—病原生物学] Q782[医药卫生—基础医学]