军事医学2024，Vol.48Issue(2)：108-114,7.DOI:10.7644/j.issn.1674-9960.2024.02.005
铜绿假单胞菌成孔毒素ExlA编码基因无痕缺失突变株的构建及其基本特性研究
Construction of the pore-forming toxin gene exlA knock-out mutant of Pseudomonas aeruginosa and its basic characteristics
摘要
Abstract
Objective To construct a non-trace deletion mutant of exlA in Pseudomonas aeruginosa strain NY8755(NY8755ΔexlA)and investigate the basic characteristics of pore-forming toxin ExlA.Methods The NY8755ΔexlA was constructed using the secondary homologous recombination method.C57BL/6J female mice ages 6 to 8 weeks were infected with NY8755 and NY8755 ΔexlA via aerosolized intratraheal inoculation respectively.Within 7 days of infection,the survival and weight changes of the mice were observed and recorded before the proinflammatory cytokines in the bronchoal-veolar lavage fluid(BALF)of the infected mice in the two groups were detected.Results The sequencing results showed that NY8755 ΔexlA was constructed.After 1×107 CFU NY8755 and NY8755 ΔexlA were infected,all the mice in the wild-type strain group died within 48 hours,while those in the mutant strain group began to die after 48 hours,and 40%of them remained alive 7 days later.The weight of surviving mice in the mutant strain group decreased but recovered gradually.After 12 hours of infection,there were more bloody exudates(redder in color)in the BALF of the wild-type strain group than in the mutant strain group,and the contents of proinflammatory cytokines interleukin-1β(IL-1β)and interleukin-17A (IL-17A)were significantly different. Conclusion Pseudomonas aeruginosa pore-forming toxin ExlA is the key pathogenic virulence factor of the exlA-positive Pseudomonas aeruginosa,which can significantly affect the survival status of mice and cause obvious inflammation in mice. Very little information is available on the action mechanisms of ExlA. In this study, The NY8755ΔexlA and the C57BL/6J mouse models infected with NY8755 and NY8755ΔexlA have been constructed that may be used for the investigation of pathogenesis of exlA-positive Pseudomonas aeruginosa.关键词
铜绿假单胞菌/基因敲除/成孔毒素/动物模型Key words
Pseudomonas Aeruginosa/gene knock-out/pore-forming toxin/animal model分类
医药卫生引用本文复制引用
张再青,周媛媛,胡凌飞,金秀玉,周冬生,高波,杨慧盈..铜绿假单胞菌成孔毒素ExlA编码基因无痕缺失突变株的构建及其基本特性研究[J].军事医学,2024,48(2):108-114,7.基金项目
病原微生物生物安全国家重点实验室自主研究课题(SKLPBS2207) （SKLPBS2207）
