Red重组系统在痢疾杆菌基因敲除中的应用研究

Study on Gene Knockout Using Red System in Shigella flexneri

利用λ噬菌体的Red系统在细菌中进行基因敲除的技术 ,最近发展较快 ,但多在大肠杆菌中进行。以alkA、wcaJ、yphF和dam 4个基因为例 ,分别在大肠杆菌和痢疾杆菌中利用Red系统进行基因敲除。对于大肠杆菌 ,除dam基因外 ,其余 3个基因均能被有效敲除 ,而在痢疾杆菌中只能敲除一个alkA基因。结果表明 ,为使该系统能有效地应用于痢疾杆菌和其它细菌 ,还需对该系统进行改进。

The technology of gene knockout using λ Red system in bacteria has been developed lately, but its application being limited to Esherichia coli . In order to get some experiences in other bacteria we select four genes, including alkA,wcaJ,yphF and dam, to test gene knockout using Red system in E.coli and Shigella . As a result , three genes were successfully knocked out in E.coli except dam while only alkA gene was knocked out in Shigella . It showed that the Red system should be improved if it was to be effectively used in Shigella and other bacteria.