中国人兽共患病学报Issue(3)：203-207,5.DOI:10.3969/cjz.j.issn.1002-2694.2015.03.003
C2株蓝氏贾第鞭毛虫 SU MO特异性蛋白酶基因的克隆、生物信息学分析及其催化活性区的原核表达
Cloning and bioinformatics analysis of Giardia lamblia C2 strain SENP gene and prokaryotic expression of SENP catalytic domain
摘要
Abstract
SUMOylation is a post‐translational modification involved in various cellular processes .SUMO‐specific protease (SENP) regulates SUMOylation by removing SUMO from conjugated substrates (deSUMOylation) and promoting maturation of SUMO precursor .In order to express Giardia lambia (C2 strain) SENP catalytic domain in E .coli ,the full‐length open reading frame of SENP was amplified by PCR from Giardia lamblia genome DNA .The PCR product about 1 620 bp was cloned into cloning vector pGM‐T .Sequencing result showed the sequence of SENP in C2 strain was identical with that in Gi‐ardia WB strain .Bioinformatics analysis showed that SENP protein possessed a 372 aa discontinuous ULP catalytic domain at C‐terminal .The catalytic domain of SENP was cloned into prokaryotic expression vector pET‐28a(+ ) .The recombinant vector pET‐28a(+ )‐SENPc was transformed into E .coli Rosetta(DE3) ,then the recombinant SENPc protein was expressed by IPTG induction .SDS‐PAGE and Western blot using anti‐His Tag antibody showed that the expression product of SENPc was a fusion protein with a molecular weight of 43 kD .The successful prokaryotic expression and bioinformatics analysis of Giardia lamblia SENP protein provide basis for further functional study of SENP .关键词
蓝氏贾第鞭毛虫/SUMO特异性蛋白酶/原核表达/生物信息学Key words
Giardia lamblia/SUMO-specific protease/prokaryotic expression/bioinformatics分类
医药卫生引用本文复制引用
李少东,周英斌,刘晓莉,禇晗,李思瑾,田喜凤,王洋..C2株蓝氏贾第鞭毛虫 SU MO特异性蛋白酶基因的克隆、生物信息学分析及其催化活性区的原核表达[J].中国人兽共患病学报,2015,(3):203-207,5.基金项目
国家自然科学基金（No ．31471954）和河北省青年科学基金（No ． C2012401039）联合资助Supported by the National Natural Science Foundation of China （No ．31471954） and the Hebei Province Science Foundation for Youths （）
