膜蛋白基因与抗生素环境适应性的关联研究

Correlation between Membrane Protein Gene and Environmental Adaptability of Antibiotics

为了深入研究植物乳杆菌(Lactobacillus plantarum)P-8在抗生素环境中的适应性进化机制,本研究采用基因敲除技术对膜蛋白基因的生物学功能进行了验证。首先,运用PCR技术扩增膜蛋白基因(LBP_cg0719)的同源臂序列,构建带有内部缺失LBP_cg0719基因的敲除载体;其次,采用Cre/lox基因重组系统,筛选双交换子L.plantarum P-8-A-1600-0719::lox66-P32-cat-lox71;最后,通过消除抗性基因,获得了突变株L.plantarum P-8-A-1600-0719,并对其耐药表型进行检测。结果表明,氨苄西林浓度为16μg/mL时,突变株L.plantarum P-8-A-1600-0719与抗生素菌株L.plantarum P-8-A-1600相比,浊度提高约2倍。因此,膜蛋白基因对植物乳杆菌P-8适应氨苄西林环境具有一定的调节作用。

In order to further study the adaptive evolutionary mechanism of Lactobacillus plantarum P-8 in the antibiotic environment,the biological function of membrane protein gene was verified using gene knockout technology.In this study,PCR was used to amplify the homologous arm sequence of a gene codes membrane protein(LBP_cg0719)to construct a knockout vector with internal deletion of LBP_cg0719 gene;the Cre/lox gene recombination system was used to screen the double exchanger L.plantarum P-8-A-1600-0719::lox66-P32-cat-lox71 to construct the mutant L.plantarum P-8-A-1600-0719;by eliminating the resistance gene,a mutant strain L.plantarum P-8-A-1600-0719 was obtained and its drug-resistant phenotype was tested.The results showed that compared with the antibiotic subculture strains,the turbidity of the mutant strain increased by about 2 when the mass concentration of ampicillin was 16μg/mL.Thus,the membrane protein gene plays a regulatory role in the adaptation of L.plantarum P-8 in ampicillin environment.