福氏2a志贺菌C0719 RNA基因突变体的构建  

Construction of C0719 RNA gene mutants of Shigella flexneri 2a

作　　者：李娜 王恒樑[2] 朱力[2] 

机构地区：[1]海军总医院检验科,北京400037 [2]军事医学科学院生物工程研究所,北京100071

出　　处：《国际检验医学杂志》2013年第17期2221-2223,共3页International Journal of Laboratory Medicine

摘　　要：目的构建福氏2a志贺菌301株已验证的但是功能未知的C0719基因突变体,以进行C0719RNA功能研究。方法根据福氏2a志贺菌301株基因组全序列,采用λ-Red重组系统对C0719RNA基因进行缺失,并经PCR验证;对野生株和C0719缺失突变株37℃时的生长曲线及生化反应进行比较研究;对野生株和5个sRNAs缺失突变株进行蛋白质组的比较研究;同时利用毒力侵袭模型来评价突变株毒力。结果构建了C0719缺失突变株;突变株初始生长均较慢,但最终和野生株状态一致;对构建成功的C0719缺失突变体进行生化特性分析,结果表明和野生株相比,301ΔC0719株利用棉子糖的能力比野生株强一些;对C0719缺失突变体进行了比较蛋白质组分析。结论获得了福氏2a志贺菌301株C0719基因突变体,为深入研究sRNA基因的功能奠定了基础。Objective To construct C0719deletion mutants of Shigella flexneri 2astrain 301for exploring the function of sRNAs in S.flexneri 2astrain 301.Methods According to the complete genome sequence of S.flexneri 2astrain 301,the sRNA gene was deleted by usingλ-Red recombination system,and confirmed by PCR.Growth of the mutant and wild strain at 37 ℃ were measured respectively,and some biochemical events of them were comparatively investigated.Results The sRNA deletion mutant of S.flexneri 2astrain 301were constructed.The mutant grew slower than wild strain at the initial period,and ultimately they showed the consistent growth state.We analysed the biochemistry characteristics of five deletion mutants.The capability to utilizing raffinose of C0719deletion mutant was more than wild strain.We also analysed the comparative proteomics of the C0719deletion mutan.Conclusion The C0719deletion mutant of S.flexneri 2astrain 301are obtained.This research will be helpful to study the functions of sRNA gene.

关 键 词：志贺菌 弗氏 基因 突变 聚合酶链反应 

分 类 号：R378[医药卫生—病原生物学]