用PCR扩增16SrRNA基因鉴定甲型副伤寒沙门菌细胞壁缺陷突变株The Identification of Cell Wall Deficient Mutants of Salmonella paratyphi A with 16SrRNA Gene Amplified by PCR
黄劲,王和
HUANG Jin~1,WANG He~2(1.Department of Biochemistry and Molecular Biology
摘要(Abstract)：
目的:探讨甲型副伤寒沙门菌细胞壁缺陷突变株的检测与鉴定方法。方法:提取甲型副伤寒沙门菌细胞壁缺陷突变株及其亲代细菌型的染色体DNA,根据甲型副伤寒沙门菌的16SrRNA基因保守序列设计引物,进行PCR扩增,对扩增产物进行琼脂糖凝胶电泳与分析。结果:甲型副伤寒沙门菌细胞壁缺陷突变株16SrRNA基因的PCR扩增产物16SrDNA,在230 bp处出现与其亲代细菌型一致的DNA条带和图谱。结论:甲型副伤寒沙门菌的细胞壁缺陷突变株虽然丧失了常规细菌学方法可检测的绝大多数表型特征,但采用聚合酶链反应方法检测该序列,有利于检测和鉴定不能自发返祖的细胞壁缺陷甲型副伤寒沙门菌。
Objective: To find a sensitive and effective method for the identification of cell wall deficient mutants(CWDMs) of Salmonella paratyphi A.Methods:Chromosome DNAs of CWDMs and bacterial type of S.paratyphi A were extracted and PCR amplification for 16srRNA genes was conducted with the chromsome DNAs as template and using primers designed after the conservative sequences of 16SrDNA of S.paratyphi A.The products were analyzed by agarose-gel electrophoresis.Results:DNA bands of 16SrRNA PCR products from both CWDMs and the bacterial type of S.paratyphi A were all at 230 bp on the agarose-gel. Conclusions: CWDMs derived from S.paratyphi A have lost most of their phenotypic features which are the foundation for their determination by routine bacteriologic methods.The technique of detecting the 16srRNA gene is more effective and sensitive in identifying CWDMs of S.paratyphi A of which the recognition is difficult with routine bacteriologic methods.
关键词(KeyWords)：
16SrRNA基因;沙门菌,甲型副伤寒;细胞壁;聚合酶链反应
16SrRAN gene;Salmonella paratyphi A;cell wall;polymerase chain reaction
基金项目(Foundation):
作者(Author):
黄劲,王和
HUANG Jin~1,WANG He~2(1.Department of Biochemistry and Molecular Biology
DOI: 10.19367/j.cnki.1000-2707.2006.04.013
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