抗噬菌体甲型副伤寒沙门菌突变菌的构建及其生物学特性

doi:10.13481/j.1671-587x.20150213

Journal of Jilin University Medicine Edition ›› 2015, Vol. 41 ›› Issue (02): 275-281.doi: 10.13481/j.1671-587x.20150213

Construction of anti-phage Salmonella Paratyphi A mutant strains and its biological characteristics

MAO Pujia^1,2, FENG Mengdie¹, HONG Yu³, MAO Xiaoping⁴, XU Zeyang¹, ZHAO Jihua³, YANG Hongwen³, SONG Wuzhan³, HUANG Fen¹, JING Shenrong¹, ZENG Weikun^1,2

1. Department of Medical Microbiology, School of Medical Science, Kunming University of Science and Technology, Kunming 650500, China;
2. Department of Clinical Laboratory Science, Medical Faculty, Kunming University, Kunming 650214, China;
3. Department of Nuclear Medicine, Kunming General Hospital, Chengdu Military Command, Kunming 650032, China;
4. Department of Urinary Surgery, Xiaogan General Hospital, Hubei Province, Xiaogan 432000, China

Received:2014-08-11 Online:2015-03-28 Published:2015-04-04

Abstract

Abstract:

Objective To construct three strains of anti-phage Salmonella Paratyphi A mutants by homologous recombination and knocking out genes in wild Salmonella Paratyphi A, and to analyze their biological characteristics.Methods The No.2,3,5 homology arm fragments obtained by PCR and fusion PCR were connected with pYG4 vector after digestion to construct the pYG4-2,pYG4-3,and pYG4-5 homologous recombination plasmids. Three strains of anti-phage mutants (No.2,3,5) were obtained after the plasmids were identificated by PCR sequencing and recombinated with wild Salmonella Paratyphi A. After the three mutant strains and wild Salmonella Paratyphi A were treated with different kinds and concentrations of antibiotics,different pH values and temperature and different concentrations of salt ions,respectively;the A₅₉₅ values were measured,and their sensitivities to the antibiotics,temperature,pH values and salt ions and the growth curves were analyzed.Results The results of PCR and fusion PCR amplification suggested that three homology arms were obtained. The PCR sequencing showed that the homologous recombinant plasmids pYG4-2,pYG4-3,and pYG4-5 were constructed successfully.The homologous recombination and anti-phage verification implied that three strains of anti-phage mutants were successfully gained. Three strains of anti-phage mutants gained kanamycin resistance and showed highly sensitivity to streptomycin;the No.2 mutant strain showed ampicillin resistance and the No.5 mutant strain performanced a high temperature tolerance. In addition,the growth curves of three mutants became obviously placid than that of wild Salmonella Paratyphi A,especially in the No.2 mutant.Conclusion The anti-phage mutants are successfully constructed and get some specially biological chatacteristics.

Key words: biological characteristics, phage resistance, Salmonella Paratyphi A, mutant,homologous recombination

CLC Number:

R378.22

MAO Pujia, FENG Mengdie, HONG Yu, MAO Xiaoping, XU Zeyang, ZHAO Jihua, YANG Hongwen, SONG Wuzhan, HUANG Fen, JING Shenrong, ZENG Weikun. Construction of anti-phage Salmonella Paratyphi A mutant strains and its biological characteristics[J].Journal of Jilin University Medicine Edition, 2015, 41(02): 275-281.

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Construction of anti-phage Salmonella Paratyphi A mutant strains and its biological characteristics

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