FUS1基因原核表达质粒构建与诱导表达  

Recombinant prokaryotic plasmid construction and high expression of FUS1 gene

作　　者：张宝[1] 霍霞[1] 彭琳[2] 齐宗利[1] 徐锡金[1] 李燕[1] 丘波[1] 郑良楷[1] 

机构地区：[1]汕头大学中心实验室 [2]汕头大学医学院附属肿瘤医院检验科,广东汕头515041

出　　处：《南方医科大学学报》2007年第5期638-640,共3页Journal of Southern Medical University

基　　金：国家自然科学基金(30600733);中国博士后科学基金(2005038187)~~

摘　　要：目的构建FUS1基因原核表达质粒,在大肠杆菌[Rosetta(DE3)2plys]中高效表达重组蛋白。方法采用PCR技术从人脐带来源的间充质干细胞中扩增出FUS1基因,并将其接入pET-32a(+),经PCR、测序鉴定后,转化Rosetta(DE3)2plys细菌,并用IPTG诱导表达。结果经过PCR、测序鉴定,克隆了pET32a(+)-FUS1重组子;在低浓度IPTG(25μmol/L)诱导3h,RosettaDE3可以高效地表达重组蛋白,约占细菌总蛋白的40%。结论成功地克隆到FUS1基因,并使其在原核系统中高效表达。Objective To construct the prokaryotic plasmid ofFUS1 gene for efficient FUSI expression in E.coli strain Rosetta （DE3）2plys. Methods The full-length FUSI gene was amplified by PCR from the total RNA of umbilical mesenchymal stem cells and cloned into pET-32a （＋） vector followed by identification with PCR and sequencing, The recombinant plasmid pET-32a（＋）-FUS1 was transformed into the E.coli strain Rosetta（DE3）2plys and the target protein expression was induced by IFFG. Results The plasmid pET-32a （＋）-FUS1 was obtained successfully as verified by PCR and sequence analysis. High expression of the fused FUS 1 protein was achieved after induction by low-concentration IPTG （25 μmol/L） for 3 h, and the recombinant FUSI protein accounted for 40% of the total bacterial protein of Rosetta （DE3）2plys, Conclusion The recombinant FUS 1 plasmid has been successfully cloned, which allows highly efficient FUS 1 expression in Rosetta （DE3）2 plys.

关 键 词：FUSI基因 基因表达 原核表达质粒 

分 类 号：R346[医药卫生—基础医学]