猪霍乱沙门菌抗噬菌体菌株筛选及受体基因突变位点分析

doi:10.16431/j.cnki.1671-7236.2022.11.041

Abstract

Abstract: 【Objective】 This study was aimed to screen the phage-resistant strains, and analyze the mutation sites of the receptor gene, in order to provide theoretical basis for revealing the mechanism of phage resistance mutation of Salmonella cholerae.【Method】 Salmonella cholerae CICC 21501 and its lytic phage vB_SenS_S528 (phage S528) were used as the research objects, the spontaneous phage-resistant strains were screened by fluctuation experiment, and the colony morphology was observed.The sensitivity to phage, adsorption characteristics, growth curve and sensitivity to temperature and pH were measured.The whole genome resequencing and PCR were used to verify the mutation site of the potential receptor gene.【Result】 A mutant strain resistant to phage S528 named B6-2 was successfully obtained.The strain B6-2 displayed rough edge of colony different from wild-type strain.Compared with wild bacteria, the growth curves showed no significant difference, but showed sensitivity to pH and temperature tolerance at 40 and 50 ℃. The adsorption rate of phage S528 to the phage-resistant strain was reduced by 60% (93% to host strain).The whole genome resequencing and comparative analysis showed that there were 3 mutation sites of the phage-resistant strain B6-2, 2 sites of PROKKA_04510 gene and 1 site of rfbC gene, respectively.The PCR product electrophoresis and sequencing results of the mutant gene fragment showed that 2 sites of PROKKA_04510 gene were not mutated, but the actual mutation site was located at 350 bp of rfbC gene, and the base was mutated from C to T.【Conclusion】 A phage-resistant strain B6-2 related to receptor changes was screened and it might prevent phage adsorption through base mutation in rfbC gene.

Key words: Salmonella cholerae; phage resistance; rfbC gene

CLC Number:

S859.79

LI Xiaoyu, ZHANG Huijun, CHI Xinyue, LI Gen, WANG Lili, WEI Bingdong, SUN Xiaowen, LI Jibin, XU Yongping. Screening of Bacteriophage Resistant Strains of Salmonella cholerae and Analysis of Mutation Sites in Receptor Gene[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4495-4502.

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Screening of Bacteriophage Resistant Strains of Salmonella cholerae and Analysis of Mutation Sites in Receptor Gene

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