摘要
目的为将恶性疟原虫的富组氨酸蛋白2（HRP2）应用于疟疾诊断和疫苗研究，本文原核表达、纯化及鉴定了恶性疟原虫的HRP2。方法将HRP2基因片段克隆入原核表达载体pGEX-4T-1，形成pGEX-4T-1／HRP2；诱导pGEX-4T1／HRP2转化的BL21菌，纯化表达产物并免疫BALB／C小鼠；以免疫色谱法鉴定表达产物，以免疫小鼠血清对受染红细胞（IRBC）进行Westemblot分析。结果成功构建了pGEX-4T-1／HRP2，经诱导表达出与GST融合的蛋白（约60KD），纯化后纯度达92％；HRP2特异性单抗可识别表达蛋白，免疫小鼠血清可识别IRBC中的原虫HRP2抗原。结论成功表达并纯化了HRP2蛋白，为深入研究和应用HRP2打下基础。
Aim in order to do research work on Histiding rich protein2 (HRP2) of Plasmodium falciparum yunan line for malaria diagnosis and Vaccine, HRP2 gene was prokaryoticly expressed.Methods .HRP2 gene rframent was cloned into prokaryotic expression vector pGEX-4T-1 to form pGEX-4T-1/HRP2, E. coli BL21 was transformed.After induerement the expresion product was purified and used to inununize BALB/C mice. Immunochromatographic test and Westem blot were done.Result pGEX-4T-1/HRP2 successfully constructed.After inducement,a GST fusion protein (about 60KD) was expressed,which can be recognized by HRP2 specific Meanwhile, the parasite HRP2 antien insied the infected red blood cells (IRBC) was detected by mice antisera in Westem blot.Conclusion HRP2 gene was successfully expressed,a good basis for future research was built up.
出处
《中国人兽共患病杂志》
CSCD
北大核心
2000年第2期35-38,共4页
Chinese Journal of Zoonoses
基金
全军区药卫生青年基金!ID962087
关键词
恶性疟原虫
富组氨酸蛋白2
GST融合蛋白
Plasmodium falciparum
Histidine rich protein2
Prokaryotic expression
GST fusion protein
