间日疟原虫MSP1-19基因的克隆与表达

李光友; 陈勇; 夏惠; 方强; 刘丹; 买月琴; 贺文欣

间日疟原虫MSP1-19基因的克隆与表达

Cloning and expression of Plasmodium vivax merozoite surface protein-1-19 gene

摘要

摘要: 目的: 构建间日疟原虫MSP1-19(Plasmodium vivax merozoite surface protein-1-19,Pv MSP1-19)基因克隆及表达的载体。方法: 将MSP1-19基因克隆入原核表达载体pET₂8a中,构建重组表达载体。以重组载体转化大肠埃希菌BL21,在IPTG诱导下表达重组Pv MSP1-19蛋白。采用亲合层析纯化重组蛋白,应用SDS-PAG电泳和Western blot对表达产物进行鉴定和分析。结果: 成功构建了质粒pET₂8a/Pv MSP1-19,并在大肠埃希菌中诱导可溶性表达的目的蛋白。表达的蛋白能与间日疟患者血清发生特异性结合反应。结论: 成功地原核表达出Pv MSP1-19目的蛋白,为间日疟的疫苗研制提供了实验基础。

Abstract: Objective: To construct a vector which clone and express Plasmodium vivax MSP1-19 Gene(Pv MSP1-19). Methods: The Pv MSP1-19 was cloned into the expression vector pET₂8a. The recombinant expression vector was transformed into E. coli,and the Pv MSP1-19 protein was expressed under IPTG induction. The recombinant protein was purified by affinity chromatography and the fusion protein was characterized by SDS-PAGE and Western blot. Results: The Pv MSP1-19 gene in plasmid pET₂8a was expressed in E. coli as a fusion protein. The fusion protein could be reacted specifically with Plasmodium vivax patient serum. Conclusions: The Pv MSP1-19 gene was expressed successfully in E. coli,which provides the necessary basis for preparing vaccine in human.

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