牛双芽巴贝斯虫rap-1基因的克隆与序列分析

摘要

对牛双芽巴贝斯虫(Babesia bigemina)潜在药物靶标Rap-1的基因进行克隆与序列分析.以兰州株牛双芽巴贝斯虫基因组为模板,经PCR扩增获得了rap -1基因,将该基因克隆到pGEM-T Easy Vector上,对重组质粒进行PCR扩增,对目的基因进行酶切鉴定及序列测定分析.结果表明,rap -1基因长度为846 bp,同源性分析结果显示,克隆序列与GenBank收录的巴西株牛双芽巴贝斯虫的核苷酸序列同源性为99.74％,说明兰州株牛双芽巴贝斯虫的rap-1与GenBank公布的参考基因具有高度同源性,rap-1基因具有很高的保守性.%The rap-1 gene, a potential target for drug screening was amplified using genomic DNA of Lanzhou babesiosis as template, and cloned into pGEM-T Easy Vector. The recombined plasmid was tested by PCR, enzyme digestion and gene sequencing. Results showed that the length of B. Bigemina rap-1 gene was 846 bp. The allogenic analysis revealed that the homology between cloned sequence and reference sequence in GenBank was as high as 99.74%, indicating that rap-1 gene was highly conservative.