流产布鲁氏菌bp26和omp10基因的同源性分析

Identity analysis of bp26 and omp10 genes of Brucella abortus

利用布鲁氏菌外膜蛋白bp26基因和omp10基因,对分离的布鲁氏菌和参考布鲁氏菌菌株进行克隆和序列分析,以明确分离的布鲁氏菌和参考布鲁氏菌菌株与基因库中的布鲁氏菌外膜蛋白bp26基因和omp10基因间的同源性。根据GenBank中的布鲁氏菌bp26基因和omp10基因,分别设计合成了1对特异性引物,以提取的分离株布鲁氏菌和2株参考布鲁氏菌的总DNA为模板,通过PCR技术扩增得到了bp26基因和omp10基因,回收纯化后将这2个基因分别连接到pMD18-T载体上,热激发转化受体菌大肠杆菌DH5α,并进行质粒提取、PCR鉴定和测序。结果表明,bp26基因全长为995bp,包含1个由753bp的核苷酸组成的完整开放阅读框,分离菌株的同源性为100%,与参考菌株M5、S2的同源性分别为100%和99.9%,与参考序列S19、870的同源性分别为99.9%和100%;omp10基因全长531bp,包含1个由396bp的核苷酸组成的完整开放阅读框,分离菌株的同源性为100%,与参考菌株M5、S2的同源性均为100%,与参考序列544的同源性为99.7%。证实bp26基因和omp10基因在各种型间的同源性都在99%以上,表明这2个基因在布鲁氏菌属的细菌中是高度保守的。

The aim of the present study was to analyze the identity of genes coding for Brucella outer membrane proteins BP26 and OMP10 among Brucella of testing or reference strains and GenBank strains.Two pairs of primers specific to Brucella bp26 and omp10 genes were designed and synthesized according to GenBank sequences,respectively.bp26 and omp10 genes were amplified by PCR from the genomic DNAs extracted from testing or 2 reference Brucella strains.The PCR products were cloned into the pMD18-T vector and sequenced,respectively.The constructed plasmids were transformed into Escherichia coli DH5α.The extracted plasmid DNA were confirmed by PCR and sequencing.Sequencing results showed that the bp26 gene was 995 bp in length containing a complete 753 bp open reading frame（ORF）.Sequence comparison revealed that the identity of bp26 genes in the testing strains was 100%,and the testing and the reference M5 or S2 strains was 100% or 99.9%,and the testing strains and the reference S19 or 870 sequences was 99.9% or 100%.The omp10 gene was 531 bp in length containing a complete 396 bp ORF.The identity of omp10 gene among the testing strains was 100%,the testing and the reference M5 or S2 strains were 100%,and the testing and the reference 544 sequence was 99.7%.The identity of bp26 gene and omp10 gene among the various types was more than 99%,indicating that they were highly conserved.