反向PCR扩增Ⅰ类整合酶基因侧翼序列

Objective: The flanking DNA regions of class 1 integrase were detected using reverse PCR. Methods: Genomic DNA of Escherichia coli strains from clinic samples were digested by the restriction enzymes Hind Ⅲ. With the help of T4 DNA ligase, the digested fragments will self-ligase. Reverse primers were designed to amplify the 5' and 3' flanking sequences of classⅠintegrase gene. Results: Study of clinic samples indicated that classⅠ integron was expressed in 68.1% of Escherichia coli strains. 1800bp DNA fragment can be obtained from flanking DNA regions of class 1 integrons by reverse PCR. Conclusion: Our clinic study indicated that distribution of class Ⅰintegron in Escherichia coli is prevalent. And we further identified the structure of classⅠ integron, there is RNA polymerase gene β subunits in flanking region of classⅠ integron. Our researches provide support for the gene analysis of the regulation of class Ⅰintegron and cloning research in the future.