单核细胞增生性李斯特菌fbpa基因敲除菌株的构建

 目的构建单核细胞增生性李斯特菌fbpa基因敲除菌株。方法 克隆fbpa及其上、下游基因，构建其载体质粒；通过酶切反应将上、下游基因分别重组到载体质粒中，形成同源重组质粒；同源重组质粒电转入细菌内，进行同源重组；采用PCR、Western blot鉴定敲除菌株。结果 单核细胞增生性李斯特菌fbpa基因敲除菌株基因组DNA无fbpa基因片段，且无FbpA蛋白表达。结论 成功构建单核细胞增生性李斯特菌fbpa基因敲除菌株

Construction of fbpA-deletion Mutant of Listeria Monocytogenes

Objective To construct the an fbpA-deletion mutant of Listeria monocytogenes. Methods The fbpA gene and it’s its upstream, downstream genes of Listeria monocytogenes was were cloned into plasmid pCRⅡ. The upstream and downstream fragments were ligated into the pAULA using restriction enzyme resulting in plasmid namedas pAULA-ΔfbpA by restriction enzyme respectively. To achieve allelic exchange, pAULA-ΔfbpA was introduced electroporated into Listeria monocytogenes by electroporation. This The mutant was confirmed by PCR and western blot. Results The fbpA gene was not screened detected in genome of fbpA-deletion mutant of Listeria monocytogenes, and FbpA did was not expressed in fbpA-deletion mutant of Listeria monocytogenes. Conclusion The fbpA-deletion mutant of Listeria monocytogenes was constructed successfully.