阪崎肠杆菌随机突变体库的构建

Generation of a Cronobacter (Enterobacter sakazakii) Mutant Library

摘要:

阪崎肠杆菌（Cronobacter（Enterobacter sakazakii））是一种以婴幼儿奶粉为主要致病源的食源性致病菌。至今人们对其功能基因所知不多，毒力因子和致病机制研究都未能找到突破口。本研究利用质粒pTnMod-okm和pBSL180构建阪崎肠杆菌随机突变体库，最终以工程菌Escherichia coli WM3064 及其所含质粒pBSL180成功构建了阪崎肠杆菌随机突变体库，并通过阪崎肠杆菌显色培养技术和抗生素抗性标记（卡那霉素50 μg/mL）对重组子进行筛选，建立了该菌功能基因组研究平台。在后继研究中不但可以实现对单个基因进行定位及功能研究还可找到有相似功能的一类基因，为抗性机制和致病机理研究奠定基础。

Abstract:

Cronobacter (Enterobacter sakazakii), a pathogen commonly found in powdered infant formula (PIF), is an important food-borne pathogen. At present, very limited information is available regarding the function of genes, virulence factors and the pathogenesis of Cronobacter. In this study, plasmid pTnMod-okm and pBSL180 were used to build Cronobacter random mutant library. Finally, the mutant library of Cronobacter was built based on transposon that contained in pBSL180 plasmid of Escherichia coli WM3064 through Chromogenic technology and antibiotic resistance labeling (kanamycin 50 μg/mL). In subsequence research, gene assignment and finding the similar function gene can be achieve, which layed the foundation for further studying the resistance mechanism and the pathogenesis of Cronobacter.