Objective:Construction of Burkholderia pseudomallei(B.pseudomallei)sRNA knockout strains and observation of their biological function.Methods:Design 9sF/9sR,9xF/9xR and R1/F1 primers,which were used to amplify the hom...Objective:Construction of Burkholderia pseudomallei(B.pseudomallei)sRNA knockout strains and observation of their biological function.Methods:Design 9sF/9sR,9xF/9xR and R1/F1 primers,which were used to amplify the homologous arm fragment upstream and downstream of the sRNA gene,through enzyme cutting,ligation,and transformation,the sRNA gene was knocked out from the B.pseudomallei by homologous recombination method.Results:The sRNA mutant was successfully constructed.In comparison with wild strain HNBP001,the growth rate,motility and biofilm formation ofΔsRNA decreased,but the antibiotic sensitivity has no differences.Conclusion:The sRNA knockout strain of B.pseudomallei was successfully constructed,laying a foundation for further research on its mechanism of regulating B.pseudomallei.展开更多
基金National Natural Science Foundation of China(No.81960002)。
文摘Objective:Construction of Burkholderia pseudomallei(B.pseudomallei)sRNA knockout strains and observation of their biological function.Methods:Design 9sF/9sR,9xF/9xR and R1/F1 primers,which were used to amplify the homologous arm fragment upstream and downstream of the sRNA gene,through enzyme cutting,ligation,and transformation,the sRNA gene was knocked out from the B.pseudomallei by homologous recombination method.Results:The sRNA mutant was successfully constructed.In comparison with wild strain HNBP001,the growth rate,motility and biofilm formation ofΔsRNA decreased,but the antibiotic sensitivity has no differences.Conclusion:The sRNA knockout strain of B.pseudomallei was successfully constructed,laying a foundation for further research on its mechanism of regulating B.pseudomallei.
