铜绿假单胞菌lasI,rhlI基因功能缺陷株的构建和生物学功能验证

构建铜绿假单胞菌lasI,rhlI基因功能缺陷株,为进一步阐明氦氧饱和高气压暴露条件诱导lasI,rhlI基因介导铜绿假单胞菌毒力调节的分子机制研究奠定基础。用双亲株接合转移法删除lasI,rhlI基因ORF编码区,通过RT-PCR方法验证目标基因编码序列mRNA的缺失;通过对细菌生长增殖能力、弹性蛋白酶代谢活性和细菌绿脓菌素分泌能力等表型的测定,验证目标基因编码序列缺失后的基因调节功能的缺陷。结果表明成功构建铜绿假单胞菌lasI,rhlI基因功能缺陷株,可作为进一步研究的基因工程菌。

Construction and characterization the lasI,rhlI functional deficit Pseudomonas aeruginosa PAO1 strains

The lasl, rhlI functional deficit Pseudomonas aeruginosa PAO1 strains were constructed for further studing the effect of hyperbaric oxyhelium exposure environment on virulence adaptation of P. aeruginosa. The bi-parental mating was used to deletion lasl, rhU gene ORF. The successful deletion of genes ORF was verified by RT-PCR and the target genes functional deficit was confirmed by detection the ability of proliferation, the activity of elastase and the secretion of pyoeyanin of different strains. The results proved the lasl, rhlI functional deficit Pseudomonas aeruginosa PAO1 strains were constructed successfully in this study.