山东医药2012，Vol.52Issue(11)：12-14,3.
抑癌基因FATS真核表达载体的构建及其定点突变
Preparation and site-directed mutagenesis of anti-oncogene FATS-expressing vector
摘要
Abstract
Objective To construct the eukaryotic expression vector and the site-directedmutagenesis vector of human tumor suppressor gene FATS for the further functional study. Methods FATS cDNA was amplified by RT-PCR from mouse testis and subsequently cloned into the eukaryotic vector p3 × Flag-myc-CMV-26 to generate p3 × Flag-FATS. Flag-FATS-C213G was constructed through site-directed mutagenesis. After transfection of p3 × Flag-FATS and Flag-FATS-C213G into U2OS cells, the expression of FATS proteins was detected by Western blot. Results The vector p3 × Flag-FATS checked by EcoR I /Kpn I digestion showed two size corresponded bands, and no other miscellaneous bands were found; DNA sequencing showed that AAU, which encoded the C213 of FATS, was changed to GAU; there were no mutation sites in other codons of FATS in Flag-FATS-C213G recombinant plasmid; the expression of FATS protein and flag tag was found in U2OS cells after transfeetion of p3 x Flag-FATS and Flag-FATS-C213G. Conclusions The eukaryotic expression vectors for FATS and its mutant can be successfully constructed, which may pave the way for deeply study on tumor suppressor function of FATS and its relationship with the risk of tumorigenesis.关键词
脆性位点相关的肿瘤抑制基因/定点突变/真核表达Key words
fragile-site associated tumor suppressor gene/ site-directed mutagenesis/ eukaryotic expression vector分类
医药卫生引用本文复制引用
闫双双,马克,仇丽,田寅,李政..抑癌基因FATS真核表达载体的构建及其定点突变[J].山东医药,2012,52(11):12-14,3.基金项目
国家重点基础研究发展计划(973计划)资助项目(2009CB526407). （973计划）
