SAFB1蛋白的表达纯化及其对乳腺癌细胞抑制作用的验证  

Expression of SAFB1 protein and validation of its breast tumor suppression

作　　者：季雯娟[1] 张磊[1] 王慧娟[1] 刘挺[1] 柏庆然[1] 吴传芳[1] 

机构地区：[1]四川大学生命科学院功能基因组实验室,成都610064

出　　处：《四川大学学报（自然科学版）》2010年第4期906-910,共5页Journal of Sichuan University(Natural Science Edition)

基　　金：国家自然科学基金重大研究计划(90919005);科技部科技重大专项(2009ZX09103-698)

摘　　要：利用RT-PCR方法从7721细胞系中分离野生型SAFB1基因,将该基因分别克隆入PET28a和pcDNA3.1载体来构建原核表达及真核表达载体.重组质粒经IPTG诱导表达后亲和层析纯化,用Western印迹实验验证蛋白表达的正确性.将SAFB1基因转入MCF7中,以RT-PCR的方法检测蛋白的表达,检测其对XOR基因的调控作用及做生长曲线验证其对乳腺癌细胞抑制作用.结果证实了SAFB1对乳腺癌细胞生长抑制作用非常明显,且对XOR基因的表达有一定的抑制作用.SAFB1 gene was cloned from 7721 cell line, and ligated into prokaryotic and eukaryotic expression vectors PET28a and pcDNA 3.1. To obtain recombinant SAFB1 protein, IPTG was used to induce its expression, followed by affinity purification. The identity of SAFB1 was confirmed by Western blot analysis. Ectopic expression of SAFB1 in MCF7 cells was measured by RT-PCR. When overexpressed, SAFB1 reduced the transcript level of hXOR, and slowed down the cell growth.

关 键 词：SAFB1蛋白 肿瘤抑制 生长曲线 hXOR基因 表达纯化 

分 类 号：Q78[生物学—分子生物学]