Abstract

Transposons are widely used in genetic engineering research and play an essential role in microbial drug resistance research. Transposase is the key to constructing efficient transposons, but no studies have reported transposons in Burkholderia pseudomallei. In this study, a new transposase gene was cloned and identified from HNBp001 isolated from Hainan province. Blast and phylogenetic tree analysis showed that the gene had high homology with IS21 transposase of Burkholderia strains from other regions. The transposase gene deletion and over-expression strains of HNBp001 were successfully constructed. The biofilm detection result shows that the over-expression strain's biofilm production was higher than the other two strains. Although there was no significant difference in the amount of biofilm production, the biofilm synthesis rate of the over-expression strain was significantly faster. Wild and deletion strains' results were the same in drug resistance, while over-expressed strains changed compared with the other two strains. Sodium Dodecyl Sulfate Poly Acrylamide Gel Electrophoresis (SDS-PAGE) revealed that there was no difference whether the transposase gene was knocked out or not. Two different sizes of proteins of over-expressed strain were significantly lower than that in the normal and knock-out strain. These results indicate that the cloned transposase genes play an essential role in the biofilm formation and drug resistance in HNBp001, but the specific mechanism remains to be further studied.

Key words: Burkholderia pseudomallei, transposase, biofilm, drug-resistance.