新疆奶牛乳腺炎致病性金黄色葡萄球菌FnbpA基因的克隆与序列分析

采集新疆奶牛隐性乳腺炎乳样,经分离、鉴定,从中得到金黄色葡萄球菌,参考GenBank中发表的FnbpA基因序列,用Oligo 6.0软件设计一对特异性引物。提取分离株中的基因组DNA,经PCR扩增得到1 654 bp的目的片段FnbpA,将其插入PMD18-T克隆载体中构建PMD18-FnbpA重组质粒,将其转入到大肠杆菌DH5α中,并进行测序分析。结果表明,克隆的FnbpA基因片段全长1 654 bp,共编码314个氨基酸。分析表明,FnbpA蛋白基因潜在的蛋白质抗原决定簇有17个,抗原性很强,与16株FnbpA基因标准序列的同源性为77.1%~100%,本研究所克隆的FnbpA基因与AM990992株亲缘性最近。

Clone and Sequence Analysis of FnbpA Gene of Staphylococcus aureus Isolated from Xinjiang Dairy Cow with Subclinical Mastitis

Staphylococcus aureus which cause subclinical mastitis in cow was isolated from Xinjiang cows and identified.According to the FnbpA gene sequences published by the GenBank,a pair of specific primers designed by Oligo 6.0.FnbpA gene was amplified by PCR with genomic DNA and inserted into pMD18-T vector which was transformed into the competent E.coli DH5α and was analyzed by sequence.The result showed that the FnbpA gene fragment is 1 654 bp which encoded 314 Amino acids.There are 17 Potential protein antigenic determinants in the predicted FnbpA gene and the antigenicity is significant.Compared with other published gene sequences,the homology of gene standard is 77.1%~100%.There are the closest relations between gene cloned FnbpA and AM990992.