摘要
为了进行抗蜱及蜱传病疫苗的研究,本试验根据GenBank中的镰形扇头蜱组胺结合蛋白(Rhipicephalus haemaphysaloideshistamine bindingprotein,RhHBP)基因cDNA序列,设计一对特异性引物(内含EcoRI/XhoI酶切位点),经RT-PCR扩增了镰形扇头蜱组胺结合蛋白RhHBP的cD-NA,大小为603 bp,将其克隆到pGEM-Teasy中并测序。在去除编码RhHBP信号肽的核苷酸序列后,将其亚克隆到pGEX-4T-1中,构建重组质粒pGEX-4T-1-RhHBP,将该重组质粒转化大肠杆菌BL21,经终浓度为1 mmol/L IPTG诱导其表达,表达的融合蛋白大小为49 ku,以包涵体的形式存在。Western Blot分析表明,该蛋白具有良好的免疫原性。
According to the Rhipicephalus haemaphysaloides histamine binding protein （RhHBP） gene sequence available in GenBank, the specific primers with EcoRI/XhoI site were designed by DNAMAN software. The RhHBP gene fragment was amplified by RT-PCR from genomic RNA and cloned into pGEM-T easy plasmid vector for sequencing. The RhHBP gene is 603bp in length. The gene fragment without the signal peptide was subctoned into vector pGEX-4T-1, and expressed in E. coli BL21 with the induction of lmmol/L IPTG. The expressed protein had a molecular weight of 49 ku. The Western blot analysis indicated that the protein could react with the serum to RhHBP.
出处
《畜牧与兽医》
北大核心
2009年第2期29-32,共4页
Animal Husbandry & Veterinary Medicine
关键词
镰形扇头蜱
组胺结合蛋白
克隆
重组表达
Rhipicephalus haemaphysaloides
histamine binding protein
clonning
recombinant expression
