恶性疟合成多肽抗原基因在大肠杆菌中表达产物的分离纯化  

Isolation and Purification of the Fused Protein Encoded by Synthetic Antigen Gene of Plasmodium Falciparum and Its Expression in Escherichia coli

作　　者：陈仕荣[1] 王昌才[1] 钟雄林[1] 胡亚芳[1] 王启松[2] 

机构地区：[1]第一军医大学生物化学教研室,广州510515 [2]上海复旦大学遗传所

出　　处：《生物工程学报》1995年第2期196-199,共4页Chinese Journal of Biotechnology

摘　　要：我们用化学方法合成的两个恶性疟原虫杂合抗原基因,即A基因和B基因,分别与表达载体pWR450·1重组并转化到大肠杆菌使其表达。经筛选获得了pWR/A·7和pWR/B·164两个表达较高的克隆菌(经扫描测定表达融合蛋白量分别为菌体总蛋白的8.8％和11.0％)。又将A基因和B基因串连后与pWR450·1重组并转化到大肠杆菌JM103株,经筛选鉴定获得了上述两个基因串连的pWR/AB·20克隆菌(表达融合蛋白量为35.8％)。为了进一步研究这3个克隆菌表达蛋白的免疫功能等活性,用8.0mol/L脲处理包涵体,离心沉淀上清液进行DEAE-sephacel柱层析,用Tris梯度缓冲液洗脱,但分离效果不好,而应用PAGE方法进行分离纯化,则可得到电泳纯、稳定又具有免疫原性的产物。The fusion protein from synthesized plasmodium falciparum hybrid antigen gene expressed in E. colt was purified. The recombinants (pWR/A ?7,pWR/B ?164, pWR/AB ?20)grew in Luria broth medium with lactose. The bacteria were harvested by centrifugation and the pelleted cells were, suspended in lysis buffer containing NP-40 and lysozyme and were treated by sonication and centrifugation. The fusion protein was isolated with SDS-PAGE and recovered by electrophoresis. One to four mg pure fusion protein can be obtained from 80-100 ml bacterial culture media. (Rabbits were immunized by the purified fusion protein and serologic assay has demonstrated with immuno-geneicity).

关 键 词：疟疾 疟原虫 合成多肽 抗原 表达产物 提纯 

分 类 号：R531.303[医药卫生—内科学] R394.8[医药卫生—临床医学]