Screening of new virulence genes of high virulence Klebsiella pneumoniae by Tn-seq
10.3760/cma.j.cn112309-20210330-00101
- VernacularTitle:Tn-seq法筛选高毒力肺炎克雷伯菌新毒力基因
- Author:
Ting ZHANG
1
;
Jimin SHI
;
Jialong CHEN
;
Gang LI
Author Information
1. 复旦大学附属金山医院检验科，上海 201508
- Keywords:
Transposon mutation library;
Serum bactericidal test;
Tn-seq;
hvKp;
New virulence genes
- From:
Chinese Journal of Microbiology and Immunology
2021;41(7):501-506
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To construct a transposon mutation library and screen new virulence genes of hypervirulent Klebsiella pneumoniae(hvKp). Methods:The transposon mutation library was constructed and treated with human serum. The changes in the abundance of the genes of library mutant strains were analyzed by Transposon sequencing (Tn-seq). Besides, KEGG (kyoto encyclopedia of genes and genomes) annotation and enrichment analysis were performed on the screened genes.Results:A total of 405 genes were screened out according to the abundance of the genes in the library treated with human serum was 20% lower than that without treated, and 351 genes, 86.7% of these genes were conserved in HS11286, NJST258_1, NTUH-K2044 and RJF293. Ten genes existed in strains NTUH-K2044 and RJF293 with high virulence, while these genes were absent in HS11286 and NJST258_1 with low virulence. The mutants with genes such as glycosyl transferase gene wzy, aggregator protein gene wzi and capsule transporter gene wza, which belong to the capsule polysaccharide gene clusters, could not be detected after serum treatment. The abundance of iron carriers gene clusters such as aerobacterin and salmonellin in each library changed less than one time. KEGG annotation results showed that most annotated genes were involved in amino acid metabolism, cofactor and vitamin metabolism, carbohydrate metabolism, etc. Conclusions:Tn-seq is a reliable method to screen functional genes. In this study, 405 candidate virulence genes of hvKp were successfully screened out, providing an experimental basis for further research on the function and regulation mechanism of new virulence genes of hvKp.
