No-scar construction and its intracellular proliferation of sifA gene-deleted Salmonella Enteritidis
LIU Gui-feng, YANG Jia-xing, DING Wei, WANG Yao-nan, CAI Yuan, ZHANG Jian, WANG Yang, GENG Shi-zhong, PAN Zhi-ming, JIAO Xin-an
Key Laboratory of Prevention and Control of Biological Hazard Factors (Animal Origin) for Agrifood Safety and Quality, Ministry of Agriculture of China/Key Laboratory of Zoonoses of Jiangsu Province/ Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou University, Yangzhou 225009, China
Abstract:Salmonella spp. is one of facultative intracellular pathogens. SCV provides a safe environment for intracellular Salmonella, enabling it to survive in macrophages without being cleared. The sifA gene involves information of SCV and promotes the survival and replication of intracellular Salmonella. We aimed to study the effect of sifA gene on intracellular proliferation of Salmonella Enteritidis and further to explore its pathogenic mechanism. The C50041ΔsifA were constructed by homologous recombination, it was verified by blue-white clones, antibiotic resistances and PCR. And assays of growth characteristics, biochemical characteristics, intracellular proliferation of C50041ΔsifA and sifA gene expression were performed. C50041ΔsifA was displayed that there was no difference in biochemical characteristics and growth speed, but its ability of survival and proliferation of C50041ΔsifA decreased in macrophages RAW264.7 compared to that of parental C50041. Whether inside or outside of macrophages RAW264.7, mRNA of sifA gene increased in the spiC gene mutant by qPCR. It was concluded that Salmonella Enteritidis C50041ΔsifA was successfully constructed, its proliferation could significantly decrease in macrophages and sifA gene expression might be regulated by spiC gene, which would lay an important academic foundation for in-depth research on proliferation in macrophages and the function of the sifA gene.
刘桂凤,杨家兴,丁炜,王耀楠,蔡媛,张健,王洋,耿士忠,潘志明,焦新安. 肠炎沙门菌sifA基因缺失株的无痕构建及其在巨噬细胞内的增殖特性研究[J]. 中国人兽共患病学报, 2020, 36(3): 178-186.
LIU Gui-feng, YANG Jia-xing, DING Wei, WANG Yao-nan, CAI Yuan, ZHANG Jian, WANG Yang, GENG Shi-zhong, PAN Zhi-ming, JIAO Xin-an. No-scar construction and its intracellular proliferation of sifA gene-deleted Salmonella Enteritidis. Chinese Journal of Zoonoses, 2020, 36(3): 178-186.