P47 is a protein expressed only in female gametocyte and located on the surface of female gametes of malaria parasites. In present study, the complete sequence of Plasmodium yoelii Pys47 was amplified by PCR and cloned into eukaryotic expression vector pcDNA3. 1＋. Cene sequencing and restriction enzyme digestion confirmed the successful construction of Pys47 plasmid. The transmission blocking effect of immune sera from BALB/c mice vaccinated with the DNA plasmid Pys47 was determined by IFA, but the anti-sera couldn't markedly inhibit the development of the protozoa from gametocyte into zygotes and ookinetes when the parasites in sexual stage were cultured with anti-sera in vitro. The results of present study implied that the specific surface protein Pys47 may not be an ideal candidate of immunizing antigen for transmission-blocking vaccine.%P47是特异性表达于疟原虫雌性配子体以及雌性配子表面的蛋白,为了评价其作为传播阻断疫苗候选抗原的免疫效果及其效应,本研究将约氏疟原虫Pys47完整编码区基因序列克隆入真核表达载体pcDNA3.1+,获得核酸质粒,肌肉注射BALB/c小鼠.通过ELISA方法检测免疫小鼠血清滴度,IFA检测疫苗的免疫活性以及免疫血清对有性阶段虫体发育的阻断效应.测序结果和酶切鉴定证实构建的质粒与要求相符;ELISA结果显示免疫小鼠血清特异性抗体滴度明显高于对照组;IFA结果显示免疫血清未能明显阻断配子体向合子与动合子的发育.因此,本实验提示作为疟原虫有性阶段特异性表达蛋白P47,可能不是一个理想的疟原虫传播阻断疫苗候选抗原.
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