突变SOD1cDNA亚克隆入酵母穿梭表达质粒pGBKT7中  

Subcloning Mutant SOD1cDNA into pGBKT7 of Yeast Expression Plasmid

作　　者：陈桂生[1] 史树贵[1] 李露斯[1] 陈康宁[1] 袁顺宗[2] 彭旭[2] 吴军[2] 罗高兴[2] 

机构地区：[1]第三军医大学西南医院神经内科,重庆400038 [2]第三军医大学西南医院烧伤研究所,重庆400038

出　　处：《中国药业》2008年第1期3-4,共2页China Pharmaceuticals

基　　金：国家自然科学基金资助项目;项目编号:30300116

摘　　要：目的将突变SOD1cDNA亚克隆入酵母穿梭表达质粒pGBKT7中。方法利用PCR方法从重组子pEGFP-N2-mSOD1中扩增突变的SOD1cDNA片断。以EcoRⅠ/SalⅠ双酶切突变SOD1cDNA片断和酵母穿梭表达质粒pGBKT7,在T4 DNA连接酶的作用下,连接突变SOD1cDNA片断和酵母穿梭表达质粒pGBKT7。转化感受态DH5α,筛选重组子,进行双酶切和测序鉴定。结果经酶切和测序鉴定,突变SOD1cDNA成功地亚克隆入酵母穿梭表达质粒pGBKT7中。结论亚克隆成功,可以对目的基因进行下一步研究。Objective To subelone mutant SODleDNA into pGBKT7 of yeast expression plasmid. Methods The SODleDNA fragment was gotten from pEGFP- N2- mSODleDNA by polymer ease reaction （PCR）. After digestion by Sal I and EeoR Ⅰ,mSOD1 fragment was subeoloned into pGBKT7 of yeast two hybird expression plasmid by action of T4 ligase. To transform competence DH5α, screening reeon by double enzyme cutting and sequencing. Results The mSOD1 fragment was subeoloned into pGBKT7 of yeast two hybird expression plasmid by identification of double enzyme cutting and sequenein. Conclusion To succeed subeloning is helpful to the next research of mutant SOD1.

关 键 词：突变SOD1cDNA 亚克隆 pGBKT7 

分 类 号：Q785[生物学—分子生物学]