BAI GUANG-XING, SUN ZHI-WEI, YU WEI-YUAN, et al. Effects of co-expression of protein folding modulators on activity of urokinase expressed in E. coli. [J]. Journal of army medical university, 2009, 31(17): 1664-1667.
BAI GUANG-XING, SUN ZHI-WEI, YU WEI-YUAN, et al. Effects of co-expression of protein folding modulators on activity of urokinase expressed in E. coli. [J]. Journal of army medical university, 2009, 31(17): 1664-1667.DOI:
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