食品科学 ›› 2017, Vol. 38 ›› Issue (4): 113-117.doi: 10.7506/spkx1002-6630-201704019

• 生物工程 • 上一篇    下一篇

单增李斯特菌srtA基因敲除菌株的构建及其生物学特性

李 森,热依汉古丽·麦麦提力   

  1. 上海理工大学医疗器械与食品学院,上海 200093
  • 出版日期:2017-02-25 发布日期:2017-02-28
  • 基金资助:
    上海理工大学博士启动基金资助项目(1000308001);上海市高校青年教师资助计划项目(10-15-308-802)

Construction and Biological Properties of srtA Gene Knockout Mutant Strains of Listeria monocytogenes

LI Sen, Zejhanguri·MAMATIRI   

  1. School of Medical Instrument and Food Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China
  • Online:2017-02-25 Published:2017-02-28

摘要: 单核细胞增生性李斯特菌是常见的食源性致病菌,其细胞壁上存在的表面蛋白与其致病性和细菌菌膜的形成密切相关,而srtA基因是介导表面蛋白膜表面定位的关键因子,通过识别特定的蛋白序列将表面蛋白共价结合在细胞壁上,也是单增李斯特菌的重要毒力基因。为深入研究srtA的功能及其对细菌毒力的调控,本研究通过同源重组技术敲除了单增李斯特菌中的srtA基因,并对基因敲除菌株的生物学特性进行了初步研究。通过生长曲线的测定,发现srtA基因敲除株的生长活性低于野生型菌株。进一步利用该菌株对星形胶质细胞系U251的侵袭实验发现,srtA基因敲除菌株的侵袭效率低于野生菌株,提示srtA基因在调控单增李斯特菌的侵袭能力方面发挥重要作用。本研究可为单增李斯特菌毒力基因调控和细菌菌膜的研究提供理论依据。

关键词: 单增李斯特菌, srtA基因, 同源重组, 生长曲线, 侵袭

Abstract: Listeria monocytogenes is a common foodborne pathogen. Cell wall-anchored surface proteins are closely related to the pathogenicity and biofilm formation of the bacterium. The srtA gene is a key factor that mediates the membrane anchoring of surface proteins, and as an important virulence gene of L. monocytogenes it can covalently bind surface proteins to the cell wall by recognizing specific peptide sequences. For a better understanding of the function of srtA and its role in bacterial virulence regulation, we knocked out the srtA gene in L. monocytogenes by homologous recombination in this study, and evaluated the biological properties of the mutant strain. Based on its growth curve examination, we found that the growth rate of the mutant strain was slower than that of the wild-type one. Furthermore, the invasion efficiency of the mutant strain in glioma U251 cells was lower than that of the wild-type one. These results indicate that srtA may play an important role in regulating the invasiveness of L. monocytogenes, which will provide a theoretical basis for further understanding of the regulatory mechanisms of virulence genes in L. monocytogenes and biofilm formation.

Key words: Listeria monocytogenes, srtA gene, homologous recombination, growth curve, invasion

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