摘要： 为明确试验布鲁菌和参考布鲁菌菌株与基因库中布鲁菌外膜蛋白BP2和OMP10基因间的同源性.利用布鲁菌外膜蛋白bp2和omp10基因,针对试验布鲁菌和参考布鲁菌菌株进行克隆和序列分析.根据GenBank发表的布鲁菌bp2基因和omp10基因,分别设计合成一对特异性引物,以提取的试验布鲁菌和2株参考布鲁菌的总DNA为模板,通过聚合酶链式反应(PCR)技术扩增得到bp2和omp10基因,回收纯化后将这两个基因分别连接到pMD18-T载体上,热激发转化受体菌大肠杆菌DH5a,质粒提取、PCR鉴定、测序.结果表明,bp2全长为995 bp,包含一个由753 bp组成的完整开放阅读框,试验菌株的同源性为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全长531 bp,包含一个由39 bp组成的完整开放阅读框,试验菌株的同源性为100%,与参考菌株M5、S2的同源性均分别为100%,与参考序列544的同源性分别为99.7%.证实bp2基因和omp10基因在各种型间的同源性都在99%以上,表明这两个基因在布鲁菌属是高度保守的.

关键词: 布鲁菌, bp2, omp10, 同源, 分析

Abstract: To clear genes homology on gene of the Brucella outer membrane protein BP26 and OMPl0 genes a- mong Brucella of testing or reference and Genbank strains,clone and Comparison for their DNA sequences.We de. signed and synthesized a specific primers with Brucella 6p26 and omplO in Genbank.The genomic DNA of bp26 and omplO of testing or reference Brucella strains was extracted from each sample PCR amplification was then carried out and thus obtaining 6p26 and omplO.The PCR produce was inserted into pMDl8-T vector and sequenced.The plasmids were transformed into E.coli DH5a.Extracted E.coli plasmid DNA,PCR detected and sequenced.Sequencing showed that bp26 was 995 bp in length,including a753 bp open reading of the complete framework.Sequence comparison revealed that homology of bp26 among the testing strains was 100%,homology of bp26 of the testing and the reference M5 or S2 strains were 100%or 99.9%,homology of 6p26 of the testing strains and the reference S19 or 870 se. quences Was 99.9%or 100%.omplO Was 531 bp containing a396 bp open reading of the complete framework.homol- ogy of omplO among the testing strains was 100%homology of omplO of the testing and the reference M5 or S2 strains were 100%.homology of omplO of the testing and the reference 544 sequence were 997%There arebp26 and omplO gene homology between the various types in more than 99%.They is highly remain stable.

Key words: Brucella, bp26, omp10, Hanology, Analysis

中图分类号: 

• S852.1

• S851.34

• S855.1+2