Cloning and bioinformatics analysis of Giardia lamblia C2 strain SENP gene and prokaryotic expression of SENP catalytic domain
10.3969/cjz.j.issn.1002-2694.2015.03.003
- VernacularTitle: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
- Author:
Shaodong LI
;
Yingbin ZHOU
;
Xiaoli LIU
;
Han ZHU
;
Sijin LI
;
Xifeng TIAN
;
Yang WANG
- Publication Type:Journal Article
- Keywords:
Giardia lamblia;
SUMO-specific protease;
prokaryotic expression;
bioinformatics
- From:
Chinese Journal of Zoonoses
2015;(3):203-207
- CountryChina
- Language:Chinese
-
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 .
