霍乱弧菌含编码霍乱毒素基因的溶源性噬菌体转染检测

Transfection of Lysogenic Phage CTXΦ Encoding Cholera Toxin in Vibrio Cholerae.

[目的 ]研究霍乱毒素基因 (ctx)在霍乱弧菌O1 群埃尔托型、古典型菌株和O1 39群菌株之间的传递现象 ,了解含编码霍乱毒素基因的溶源性噬菌体 (CTXΦ)在ctx基因转移中的作用。 [方法 ]从人工构建的供体菌中诱导出带有标记的CTXΦ并转染至受体菌 ,用PCR扩增和Southern杂交等方法检测受体菌、转染子及其质粒的目的基因片段。 [结果 ]从受体菌O395、569B(古典型 )和 80 0 71 (埃尔托型 )的转染子中均检出带有供体菌标记的质粒 ,PCR和Southern杂交结果表明这些质粒是CTXΦ的复制形式。 [结论

Objective\]To study the transmission of gene ctx which encodes cholera toxin in different type of Vibrio cholerae and the role of lysogenic phage CTXΦ in transmission.\PCR and Southern blot were used to detect the recipient strains of Vibrio cholerae transfected by genetically marked derivative of the phage CTXΦ for the fragment of the target gene.\Plasmid , not present before transfection,was found in recipent strains after transfection . PCR and Southern blot hybridization assays showed that the plasmid found in classical strains O 395 ?569B and Eltor strain 80071 was the CTXΦ replicative form and CTXΦ genome was not integrated into the chromosome of recipent strains . \[Conclusions\]Gene ctx can transfer horizontally between different types of Vibrio cholerae through the vector of lysogenic phage CTXΦ.