奇异变形杆菌、洛菲氏不动杆菌16s rRNA3’-23s rRNA 5’间序列的克隆及测序

Cloning and sequencing of the 16s rRNA intergenic spacer region of proteus mirabilis and acinetobacter iwoffii

目的:对奇异变形杆菌、洛菲氏不动杆菌16s rRNA 3’~23s rRNA 5’间序列(16S^23S rRNA intergenic spacer region,ISR)进行克隆测序,为分子探针技术鉴定两菌奠定基础。方法:针对临床常见致病菌16s rRNA 3’~23s rRNA 5’间序列两端的16S及23S rRNA保守序列设计PCR扩增的通用引物,对两菌进行扩增,利用PMD-18TVector质粒对两菌的PCR产物进行T载体克隆构建,测序后申报Genbank。结果:两菌的通用引物扩增产物构建的T载体克隆,测序结果经Blast分析,确定为两菌的ISR序列,申报Genbank获得接收。结论:成功的对奇异变形杆菌、洛菲氏不动杆菌ISR序列进行了克隆测序,该序列可以用于两种菌的通用引物PCR扩增技术鉴定。

Objective：To clone and sequence of 16S～23S r RNA intergenic spacer region of Proteus mirabilis and Acinetobacter lowffii for further identification of these two bacteria by means of molecular probe. Methods： The PCR universal clinical common pathogenic bacteria primers for 16S and 23S rRNA＇s conserved sequences were designed. Then the genomes of these two bacteria were amplified,T Vector clones were constructed with PMD-18 T Vector ad PCR products. The vectors were sequenced and the sequence was submitted to Genbank, Results： T Vector clones were constructed with products amplified by using universal primers; according to Blast analyses, the results of sequencing were determined as ISR sequences of the two kinds of bacteria. And the reports to Genbank were accepted.Conclusion： Genbank accepted the cloning and sequencing results of Proteus mirabilis and Acinetobacter lwoffii as new sequences.And these sequences can be used to identify the two bacteria by universal primers.