Effects of co-expression of protein folding modulators on activity of urokinase expressed in E. coli
- VernacularTitle:共表达折叠调节因子对尿激酶功能性表达的影响
- Author:
Guangxing BAI
;
Zhiwei SUN
;
Weiyuan YU
;
Jin LI
- Publication Type:Journal Article
- Keywords:
co-expression;
chaperones;
disulfide isomerase;
urokinase;
activity
- From:Journal of Third Military Medical University
2003;0(17):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To study the coexpression of protein folding modulators,including chaperones(DnaK,DnaJ,and GroEL-GroES) and disulfide isomerase Dsb molecules(DsbA and DsbC) on the bioactivity of eukaryotic expressed urokinase(UK).Methods The gene sequences of DnaK,DnaJ,GroEL-GroES,DsbA and DsbC were isolated from the chromosome of E.coli by PCR.The obtained fragments mentioned above were cloned into the plasmid pBAD-1 under the control of araB promoter and then the plasmids were transformed into E.coli BW25113 and HB101 together with a compatible vector pQE-UK.The amount of expressed UK was measured by Bradford method and its activity was analyzed by fibrin plate method for its fibrinolysis.Results The activity of expressed UK was increased by co-expressing with DnaK,GroEL-GroES,DsbA,and DsbC in BW25113.Among them,the highest activity was achieved by co-expressing with GroEL-GroES.Conclusion Co-expressing of chaperones or disulfide isomerase can improve the bioactivity of UK expressed in E.coli.
