用新法制备K88ac基因探针

A NEW METHOD PREPARED GENE PROBE

由于遗传工程技术的发展,基因探针被广泛应用。探针的DNA大都来自于大肠杆菌的重组质粒,这样制备的探针易污染该菌的染色体和载体DNA,作菌落杂交时易出现假阳性。为了克服这些缺点,用枯草杆菌质粒作载体克隆K88ac基因的EcoRI小片段。以枯草杆菌重组质粒作探针,与含K88ac基因的菌株杂交呈阳性,而不含K88ac基因的菌株呈阴性,且阴阳性有明显区别。

Plasmid pNZ8802 containing K88ac gene was digested by EcoRI, and the small fragment was cloned to vector plasmid pUB110. One of the hybrid plasmids was named as pNZ8803 which was used to gene probe for detecting varity of strains. Strains containing K88ac or K88ab gene were all positive hybridization, and strains which did not containing K88ac gene were all negative hybridization. The result indicated that the gene probe was highly specificity and sensetivity.