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目的：体外扩增编码恶性疟原虫传播阻断抗原Pfs和Pfs48／45的基因序列，为进一步对其进行克隆和体外高效表达创造条件。方法：特定寡核苷酸引物的设计、合成与纯化；恶性疟原虫FCC1／HN株体外培养；利用碱裂解法从培养的恶性疟原虫FCC1／HN株提取染色体DNA；PCR扩增和琼脂糖凝胶电泳分析。结果：从恶性疟原虫FCC1／HN株基因组DNA中特异扩增出编码Pfs25和Pfs48／45基因序列，其片段大小分别为657bp和1，359bp。而用间日疟原虫基因组DNA为模板作对照，无扩增条带出现。结论：体外扩增编码恶性疟原虫传播阻断抗原Pfs25和Pfs48／45基因序列与预期长度相符合，从而，为该基因的克隆、测序和表达奠定基础。
OBJECTIVE: To amplify the gene sequence encoding Pfs and Pfs48 / 45, which are the transmission antigens of Plasmodium falciparum in vitro, and to create conditions for further cloning and efficient expression in vitro. METHODS: Design, synthesis and purification of specific oligonucleotide primers; in vitro culture of Plasmodium falciparum FCC1 / HN strain; extraction of chromosomal DNA from P. falciparum FCC1 / HN strain by alkaline lysis; PCR amplification and agarose coagulation Gel electrophoresis analysis. Results: The Pfs25 and Pfs48 / 45 gene sequences were amplified from the genomic DNA of Plasmodium falciparum FCC1 / HN strain. The fragment sizes were 657bp and 1,359bp, respectively. With P. vivax genomic DNA as a template for control, no amplification bands appear. CONCLUSION: In vitro amplification of the sequences encoding the Pfs25 and Pfs48 / 45 genes encoding for the transmission of the Plasmodium falciparum parasite agrees well with the expected length, thus laying the foundation for the cloning, sequencing and expression of this gene.