摘要
采用缺口填补法将恶性疟原虫有性期Pfs230和Pfs48／45抗原中的2个T细胞表位，3个B细胞表位以及人白介素-Ⅰ中的一个T细胞激活位点按预设方案，拼接成复合多价基因PfSI。复合基因全长147bP，编码43肽复合抗原，经分离、纯化后，插入质粒pcDNA3的多克隆位点，构建重组载体pcDNA3／PfSI。再将重组载体转化大肠杆菌JM109，用氨苄青霉素和PCR扩增法筛选阳性克隆，最后用限制性内切酶对阳性克隆进行鉴定。复合基因PfSI拼接、克隆的成功为在体外高效表达恶性疟原虫传播阻断抗原创造了条件。
Using filling-gap method,a hybrid gene named PfSI coding for several epitopes of Pfs230 and Pfs48/45 in the sexual stage of Plasmodium falciparum and a exogenous T cell epitope from interleukin-I(IL-I)was designed and chemically synthesized. The gene was composed of 147 base pairs and coded for a 43 amino acid peptide. After purification,the target gene PfSI was inserted into plasmid pcDNA3 at poly linker.Then,the recombinant vector pcDNA3/PfSI was transformed into E.coli JM109. Positive clones were screened by ampicillin and PCR technique,and identified with restriction endonucleases. The success of recombination of the hybrid gene PfSI will facilitate the high level expression of transmission-blocking antigens of Plasmodium falciparum.
出处
《中国人兽共患病杂志》
CSCD
北大核心
1997年第3期10-12,共3页
Chinese Journal of Zoonoses
基金
中国博士后和广东省博士后科学基金
关键词
疟原虫
恶性疟原虫
传播阻断抗原
克隆
化学拼接
Plasmodium falciparum,Gametocyte, Transmission-blocking antigen,Cloning
