Construction and characterization of the stk and stp1 knockout mutant in streptococcus suis type 2
10.16571/j.cnki.1008-8199.2018.05.002
- VernacularTitle:2型猪链球菌stk/stp1基因敲除株的构建及特性研究
- Author:
Chao-Long LI
1
;
Jian ZHANG
;
Qian-Qian WU
;
Hong-Fen HOU
;
Hui-Fang ZHANG
;
Chang-Jun WANG
;
Xiang-Rong CAO
;
Xiu-Zhen PAN
Author Information
1. 南京师范大学生命科学学院
- Keywords:
streptococcus suis type 2;
gene knockout;
biological properties;
serine /threonine kinases and phosphatases
- From:
Journal of Medical Postgraduates
2018;31(5):453-458
- CountryChina
- Language:Chinese
-
Abstract:
Objective Serine /threonine kinases (STK) and phosphatases (STP) regulate various physiological activities of prokaryotes by reversible phosphorylation of proteins .This paper aimed to study the effects of simultaneous deletion of the stk and stp1 genes on the biological characteristics and pathogenicity of streptococcus suis type 2, the Chinese virulent strain 05ZYH33. Methods The double mutant of the stk and stp1 genes of 05ZYH33 was constructed by homologous recombination .The biological characteristics of the wild strain 05ZYH33 and the mutant strain Δstk/stp1 were compared.The effects of the stk and stp1 deletion on bacterial virulence was analyzed using cell adhesion assay , anti-phagocytosis assay and the mouse model of infection . Results RT-PCR showed that the stk and stp1 genes were replaced by the spectinomycin resistance gene Spc r and the mutant strain was successfully constructed .Experi-ments of biological characterization revealed gradually increased value of 05ZYH33 and Δstk/stp1 at 2 hours after inoculation and a plateau period at 7 hours.The logarithmic phase of the mutant strain (A600≈0.4) was 1 hour later than that of the wild one , and the bacterial den-sity of the former was lower than that of the latter after the plateau pe -riod (0.8 vs 1.0).On the blood plates of 05ZYH33 and Δstk/stp1 were observed greyish, round, semitransparent, wet and smooth-sur-faced tiny bacterial colonies , around which there were hemolysis rings with no significant differences in colony morphology and hemolytic ac -tivity.In the experiment on pathogenicity , the mice of the 05ZYH33 group all died within 12 hours while 9 of the 30 mice in the Δstk/stp1 group died within 12 hours and all died within 24 hours. Conclusion The simultaneous deletion of the stk and stp1 genes may mainly affect the regulation of the proteins associated with bacte -rial proliferation and division.
