甲型副伤寒沙门氏菌噬菌体PSPA1感染宿主相关基因分析  

Analysis of Host Genes Involved in Infection of Salmonella Paratyphi A Bacteriophages PASP1

作　　者：毛普加 洪愉[2] 冯金[1] 冯梦蝶[1] 许泽仰 黄芬[1] 井申荣[1] 曾韦锟[1] 

机构地区：[1]昆明理工大学医学院,昆明市650500 [2]中国人民解放军成都军区昆明总医院核医学科,昆明市650032

出　　处：《医学分子生物学杂志》2014年第2期85-89,共5页Journal of Medical Molecular Biology

基　　金：国家自然科学基金（No.31160193）,云南省教育厅科学研究基金（No.2010Y398）,云南省应用基础研究面上项目（No.2010ZC055,2012FB135）

摘　　要：目的分析影响甲型副伤寒沙门氏菌（副甲）噬菌体（PSPA1）感染宿主相关基因。方法诱导副甲利福平抗性株（副甲Rif^＋）；将副甲Rif^＋与SM10λpir／pSC189接合转座，噬菌体PSPA1处理转座菌液后，涂布卡那霉素、利福平双抗平板筛选转座突变菌；提取抗噬菌体突变菌基因组，热不对称PCR、回收片段并测序；测序结果提交NCBI进行序列比对，确定转座质粒插入副甲基因组的位点。结果成功诱导出副甲Rif^＋；筛选到6株抗噬菌体的突变菌；测序、NCBI比对发现6个不同的插入基因。结论利用转座子插入获得抗噬菌体感染甲型副伤寒沙门氏菌突变体，对这些突变体进行PCR、测序分析，说明可能有6个基因及相应的酶或蛋白与抗噬菌体相关，为进一步分析噬菌体与寄主菌相互作用关系提供基础。Objective To analyze the related genes of Salmonella paratyphi （S. Paratyphi A bacteriophage PSPA1 in the host infection. Methods Rifampicin-resistant strains of S. Para- typhi A were induced （ S. Paratyphi A Rif^＋ ） After conjugation of S. Paratyphi A Rif＋ with SM10λpir/pSC189, the mixer was treated with phage PSPA1, and then spread on the plate contai-ning kanamycin and rifampicin to screen the mutants. The genomes of anti-phage mutants were ex-tracted as template of the thermal asymmetric interlaced PCR （Tail-PCR） . After amplification, the products were purified and sequenced. The sequence alignment was performed at NCBI to deter-mine the sites where the transposon inserted. Results The rifampicin-resistant strains of S. Paratyphi A were induced successfully. Six anti-phage mutants were screened. Among the six mutants, different inserted genes were found by sequencing and alignment. Conclusion The phage-resistant strains of S. Paratyphi A were obtained through transposon conjunction. The flan- king sequence of insertion sites of mutant strains was acquired by Tail-PCR. The sequencing results showed that the six genes, their encoded enzymes or other proteins might be involved in phage re-sistance. The present study laid a foundation for further study of relationship between phage and host.

关 键 词：甲型副伤寒沙门菌 接合转移转座 突变菌 噬菌体 

分 类 号：Q939.47[生物学—微生物学] R378.2[医药卫生—病原生物学]