间日疟原虫乳酸脱氢酶编码区全长基因的克隆、序列分析及线性B细胞表位预测  

Cloning,sequence analysis and linear B-cell epitopes prediction of full-length gene in coding region of lactate dehydrogenase from Plasmodium vivax Anhui

作　　者：方强[1] 夏惠[1] 王雪梅[1] 齐文娟[1] 常雪莲[1] 高琪[2] 

机构地区：[1]蚌埠医学院病原生物学教研室,安徽省感染与免疫重点实验室,蚌埠233030 [2]江苏省寄生虫病防治研究所,卫生部寄生虫病预防与控制技术重点实验室,无锡214064

出　　处：《中国人兽共患病学报》2010年第6期519-523,共5页Chinese Journal of Zoonoses

基　　金：国家科技重大传染病专项(2008ZX10004-011);卫生部寄生虫病预防与控制技术重点实验室开放课题(WK008-002)联合资助

摘　　要：目的克隆中国间日疟原虫乳酸脱氢酶(PvLDH)编码区全长基因,并对其进行生物信息学分析。方法自间日疟原虫抽提RNA,根据已知的PvLDH基因设计特异性引物,采用RT-PCR扩增PvLDH编码基因,将其克隆至pMD18-T载体,经PCR和双酶切鉴定后进行序列测定,利用生物信息学在线工具对序列进行分析并做B细胞表位预测。结果 PCR产物电泳结果显示所克隆的基因为951bp,基因测序结果与GenBank报道的基因序列有一个碱基差异,但编码氨基酸无差异。在线分析预测出12个B细胞表位,与恶性疟原虫乳酸脱氢酶预测表位对比分析,发现一个PvLDH特异性表位。结论成功克隆了我国间日疟原虫乳酸脱氢酶编码区全长基因,并预测出1个PvLDH特异性线性B细胞表位。The aim of this study is to clone and sequence the full-length gene in coding region of lactate dehydrogenase（LDH）from Plasmodium vivax（P.vivax）Anhui,and predict linear B-cell epitopes of P.vivax LDH（PvLDH）for the sake of providing a foundation on the development of rapid diagnosis method for vivax malaria based on P.vivax specifically monoclonal antibody.The PvLDH encoding gene from RNA of P.vivax Anhui was amplified by RT-PCR,and then the 951bp PCR-amplified product was cloned into pMD18-T vector and sequenced.The DNA and its protein sequence were analyzed and the linear B-cell epitopes of PvLDH were predicted.Sequence analysis for the cloned genes in pMD18-T vector showed one base in the full-length gene in coding region of LDH from P.vivax Anhui was different from that of P.vivax Sal-I and Belem,but there was no difference in deduced amino acid of PvLDH.A total of 12 linear B-cell epitopes of PvLDH were predicted from deduced protein sequence on line and a specific linear B-cell epitope was predicted by comparing with the linear B-cell epitopes of PfLDH.Results indicate that the PvLDH full-length gene in coding region is successfully cloned and a specific linear B-cell epitope is predicted successfully.

关 键 词：间日疟原虫 乳酸脱氢酶 克隆 序列分析 B细胞表位 

分 类 号：R282.3[医药卫生—中药学]