大鼠Sirt1基因原核表达质粒的构建和融合蛋白表达.pdfVIP

·417， 大鼠Sirtl基因原核表达质粒的构建及融合蛋白表达 耿义群傅玉才1徐小虎王伟1许锦阶1(汕头大学医学院法医学教研室，广东汕头515041) 【摘要】 easy载体后亚 长506bp，经酶切鉴定及DI、IA序列测定，证实pEr41一Sirtl重组质粒构建正确，表达融合蛋白分子量约为48kD。结论成功构建重组原核表达质粒 pET41．Sial，并能表达融合蛋白，其分子量大小与预期一致。 【关键词】大鼠；Sirtl；基因表达；重组蛋白 【文献标识码】A 【中图分类号】11394．3 【文章编号】1005-9202(2007)05-0417-03 Construction withratSift1 andfusion otprokaryotieexpressionplasmld gene proteinexpression GENG Yi-Qlm，FUYu-Cai，XIJXiao—ltu，et以 ofForensic ofShantou Department Medicine，MedicalCollege University，Shantou515041，Guangdong，China Toconstructa 【Abstractl objeetive recombinant withratSirtlcDNAandtoobtainthefusion expressionolasmid A ofratSialcDNA BamHlJXh01were RT—PCRThe w羽clonedinto vectorand fragment containing amplifiedby fra瘁nent pGEM-Teasy subelonedinto B也lcellswerelransformedtherecombinantThefusion Willsin． PE’r4lvector．Subsequently，E．eoli bv vlasmid protein．which duced andWilts SDS-PACE．ResultsA of506ofmtSirtleDNAwas RT— by∥rGw柏purified analyzedby fragmentbp amp肺edby PCRTheconstructionoftherecombinant confirmed fusion of48kDwas and plasmidPET41-Sialwasbysequencing．Theprotein expressed Therecombinanthasbeen ofratSirtl1311／1be inE．coli i)urified．Conclusions constructed．Thefusion plasmid successfully protein expressed BL2lcellswiththe toolecular expected weight_ 【KeYwords】Rat：Sial；Geneexpression；Recombinantprotein information 沉默信息调节因子-2(Sil