镰形扇头蜱P0基因的克隆表达及初步鉴定  

Cloning,expression and preliminary identification of Rhipicephalus haemaphysaloides P0 gene

作　　者：张玉婷[1,2] 龚海燕[2] 周勇志[2] 巴音查汗[1] 周金林[2] 

机构地区：[1]新疆农业大学动物医学学院,新疆乌鲁木齐830052 [2]中国农业科学院上海兽医研究所,上海200241

出　　处：《畜牧与兽医》2015年第5期12-15,共4页Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(No.31172095)

摘　　要：P0分子是一种核糖体蛋白,具有抗蜱免疫作用。本文采用RACE技术从镰形扇头蜱(Rhipicephalus haemaphysaloides)中获得了P0全长基因。通过对P0全长基因序列进行分析发现,P0全长基因为1 118 bp,具有一个编码319个氨基酸的开放阅读框,没有信号肽序列。推测P0的蛋白大小约为35 ku。P0与其他蜱种具有很高的同源性,与微小扇头蜱(Rhipicephalus microplus)和长角血蜱(Haemaphysalis longicornis)的P0基因同源性分别为94%和87%。编码区克隆到表达载体p GEX-4T-1中,转化表达宿主菌BL21,可表达出分子量约为60 ku的重组融合蛋白。表达的蛋白以包涵体的形式存在于菌体中。表达产物经GST树脂纯化后可得到纯度较高的目的蛋白。抗P0重组蛋白鼠血清可以识别蜱体内的P0蛋白,大小在35 ku。研究结果将为镰形扇头蜱新型防治技术研究提供基础。P0 is a ribosomal protein with anti-tick immunity. In this paper, the full-length gene sequence of Rhipicephalus haemaphys- aloides PO were obtained by RACE technology. The full length of the P0 gene was 1 118 bp. It has an open reading frame encoding 319 ami- no acids without signal peptide sequence. The molecular weight of P0 protein was predicted to be about 35ku. The R. haemaphysaloides PO gene has high homology with other tick species. Compared with Rhipicephalus microplus （accession number： KC845304） and Haemaphysalis longicornis （accession number： EU048401 ） , the homologies of P0 gene were 94% and 87% , respectively. The coding region was cloned into the expression vector pGEX-4T-1, and the recombinant plasmid was transformed into Escherichia coli BL2 and then expressed by indu- cing with 1 mmol/L IPTG. The results showed that the recombinant protein was about 60ku. The expression product was purified by GST res- in and identified by mouse serum against the recombinant protein. The preliminary identification of P0 protein from R. haemaphysaloides will provide a basis for development of the anti-tick vaccine.

关 键 词：镰形扇头蜱 P0基因 克隆 表达 鉴定 

分 类 号：S852.74[农业科学—基础兽医学]