摘要
以霍乱毒素Ｂ亚基（ＣＴＢ）基因为载体，构建了含不同抗原表位的恶性疟原虫的融合基因ＣＴＢ／ＡＴＥ和ＣＴＢ／ＡＷＴＥ。前者除含有恶性疟原虫裂殖子表面主要抗原表位杂合多肽基因ＳＰｆ６６外，还含有很强的Ｔ辅助细胞表位ＣＳＴ３和Ｔｃ细胞表位；后者在此基础上将我国发现的Ｂ细胞表位ＮＫＮＤＤ基因经８次串联后融合其中。两种形式的融合基因经测序正确后转入大肠杆菌ＴＫ１０４６中，产量分别为１０ｍｇ／Ｌ及５ｍｇ／Ｌ。表达产物ＣＴＢ／ＡＷＴＥ经亲和层析纯化双抗夹心ＥＬＩＳＡ测定表明，该融合蛋白在保留了与抗ＣＴＢ抗体结合的同时，与抗ＮＫＮＤＤ单抗的结合效价达１：８０００。
Two fusions genes, CTB/ATE and CTB/AWTE, which contain sequences coding for multiple antigenic epitopes of malaria Plasmodium falciparum and cholera toxin B subunit (CTB) gene used as a carrier were constructed. The former consisted of SPf66, the chimeric epitopes of merozoitic antigens, and T cell epitopes CSTE including Th determinant CST3 and Tc determinant CSP(368 390). The later was constructed by inserting a tandem repeat of eight times of a B cell epitope, NKNDD, to the former one. These two plasmids containing the correct genes under the control of Lac promoter were transferred to bacteria Escherichia coli , TK1046. The expression levels were 10mg/L for CTB/ATE and 5mg/L for CTB/AWTE. CTB/AWTE was purified on affinity chromatography column and tested by enzyme linked immune absorbent assay (ELISA). It showed that this fusion protein could bind not only with monoclonal antibody (McAb) against NKNDD at a high titre of 1:8000 but also with antibody against CTB.
出处
《生物工程学报》
CAS
CSCD
北大核心
1996年第S1期22-26,共5页
Chinese Journal of Biotechnology
基金
"863"课题资助项目
关键词
恶性疟原虫
抗原表位
霍乱毒素B亚基
融合基因
抗原性
Malaria Plasmodium falciparum , epitope, cholera toxin B subunit, fusion gene, antigenicity
