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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。
Two T cell epitopes, three B cell epitopes and one T cell activation site in human interleukin-I in the sexual stage Pfs230 and Pfs48 / 45 antigens of Plasmodium falciparum were selected according to the default protocol by gap-fill method. Spliced into a composite multivalent gene PfSI. The full-length cDNA of the complex gene was 147bP encoding 43 peptide complex antigen. After isolation and purification, the recombinant plasmid pcDNA3 / PfSI was inserted into the multiple cloning site of plasmid pcDNA3. The recombinant vector was transformed into E. coli JM109. The positive clones were selected by ampicillin and PCR amplification. Finally, the positive clones were identified by restriction enzyme. The success of the PfSI splicing and cloning of the composite gene has created the conditions for the efficient expression of the anti-Plasmodium falciparum antigens in vitro.