title: [Construction of SW480 cell model identifying Shigella virulent genes].
abstract: Signature-tagged mutagenesis (STM) is a novel technology with high throughput screening ability to identify virulent genes of pathogen in vivo. An appropriate animal or cell line model is one of prerequisites by exploiting this technique. In order to apply STM to Shigella flexneri, RC426 was constructed as an attenuated mutant with chloramphenicol resistance and aroA and virG genes inactivated by homologous recombination; another attenuated strain T32 was used as an oral S. flexneri 2a vaccine due to a spontaneous deletion in three loci (ipaBCDA, invA and virG) on the virulence plasmid. The wild type strain 2457T had the invasion ability into host cells. The three strains, RC426, T32 and 2457T, were mixed together to invade colon cancer cell line SW480, and the distinct strains were recovered and counted from cell lysates of invaded SW480 in different time. The results showed that there were statistically significant differences between the amounts of two attenuated strains recovered and that of virulent strain within 12h invasion, indicating SW480 was a suitable cell model for applying STM to screen virulent genes of Shigella flexneri.
authors.affiliation: Beijing Institute of Biotechnology, Beijing 100 071, China.
authors.forename: Xiao
authors.initials: X
authors.lastname: Yao
authors.forename: Heng-Liang
authors.initials: HL
authors.lastname: Wang
authors.forename: Zhao-Xing
authors.initials: ZX
authors.lastname: Shi
authors.forename: Xiao-Yu
authors.initials: XY
authors.lastname: Yan
authors.forename: Er-Ling
authors.initials: EL
authors.lastname: Feng
authors.forename: Quan-Hong
authors.initials: QH
authors.lastname: Liu
authors.forename: Guo-Fu
authors.initials: GF
authors.lastname: Su
authors.forename: Liu-Yu
authors.initials: LY
authors.lastname: Huang
mesh_terms: D000900:Anti-Bacterial Agents / Q000494:pharmacology; D045744:Cell Line, Tumor; D003110:Colonic Neoplasms / Q000473:pathology; D004351:Drug Resistance; D004847:Epithelial Cells / Q000166:cytology / Q000382:microbiology; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial; D005798:Genes, Bacterial; D006801:Humans; D016296:Mutagenesis; D012763:Shigella flexneri / Q000235:genetics / Q000472:pathogenicity*; D013839:Thiamphenicol / Q000031:analogs & derivatives / Q000494:pharmacology; D014774:Virulence
chemical_list: D000900:Anti-Bacterial Agents; C035534:florfenicol; D013839:Thiamphenicol