Exogenous Expression of IscR from Aeromonas veronii Regulates the Oxidative Stress Tolerance of Escherichia coli
In this study,the iscR gene was amplified using the genome of(Aeromonas veronii)as the template,double digested by enzymes,and ligated with the expression vector pET28a.The ligation products were transformed into E.coli BL21 competent cells by electroporation,and the positive transformants were selected.Finally,the recombina strain BL21-pET28a-iscR that carries the recombinant vector pET28a-iscR was obtained.Taking the recombinant strain as the research object,the expression of IscR protein was induced by adding a specific concentration of IPTG,then the strain BL21-pET28a-iscR and the control strain BL21-pET28a transferred to the no-loaded plasmid was treated by the final concentrations of H2O2 of 0,0.5,1.0,1.5 and 2.0 mmol/L for 30 min,finally the colony growth of bacteria after different H2O2 concentration treatment was observed by concentration gradient dilution point plate experiment.The results showed that,as compared with the control,the recombinant strain BL21-pET28a-iscR exhibits higher resistance to the oxidative stress under the same H2O2 final concentration treatment,suggesting that IscR protein from A.veronii can be expressed heterologous and function.Our study preliminarily proved that IscR protein of A.veronii was also involved in the regulation of bacterial response to oxidative stress,which provide the basis for further exploring the function of IscR protein and analyzing the molecular mechanism of bacterial response to oxidative stress.
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