Ⅱ型猪链球菌二元信号转导系统2148hk/rr基因敲除突变体的构建

Development of the 2148hk/rr Gene Knock-out Mutant of Streptococcus suis Type 2

摘要:

构建猪链球菌2型(Streptococcus suis type 2)强毒株05ZYH33二元信号转导系统2148hk/rr基因敲除突变体。构建中间为壮观霉素抗性基因, 两侧为2148hk/rr编码基因上、下游同源序列的基因敲除质粒, 通过同源重组筛选2148hk/rr编码基因敲除突变体。PCR分析和Southern杂交结果均显示2148hk/rr编码基因完全被壮观霉素抗性基因替代, 基因敲除突变体构建成功。筛选获得05ZYH33二元信号转导系统2148hk/rr基因敲除突变体, 为阐明该调控系统在猪链球菌致病过程中的作用奠定了基础。

关键词:猪链球菌2型, 二元信号转导系统, 基因敲除, 突变体

Abstract:

To construct gene knock-out mutant of a two-component signal transduction system named 2148hk/rr in Streptococcus suis type 2 virulent strain 05ZYH33. Recombinant gene knock-out vector was constructed consisting of Spcr cassette with flanking homology regions to the target genes, the isogenic 2148hk/rr-deficient mutant was screened by allelic replacement. PCR analysis and Southern hybridization confirmed that the coding genes of 2148hk/rr were replaced completely by spcr cassette. Conclusion The mutant of 05ZYH33 2148hk/rr system was successfully constructed, which laid the foundation for farther research on the role of this two-component signal transduction system during infection.

Key words:Streptococcus suis type 2, Two-component signal transduction system, Gene knock-out, Mutant