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目的：构建夏氏疟原虫早期滋养体的ｃＤＮＡ文库，并从中筛选出表达Ｐｃ９０的阳性克隆。方法：采用ＧＴＣ／ＣｓＣｌ法提取ＲＮＡ，ｃＤＮＡ克隆入λ－ＺＡＰ噬菌体，用抗Ｐｃ－ＨＣＰＭ血清３次筛选并经抗Ｐｃ９０单克隆抗体复筛，阳性克隆经体内剪接后行限制性内切酶酶切分析。结果：该ｃＤＮＡ文库有１．９×１０６个重组体。用抗Ｐｃ９０－ＨＣＰＭ的多克隆抗体筛选后，得到１６个阳性克隆，其长度分别为：０．６ｋｂｐ，１．４ｋｂｐ，１．６ｋｂｐ，２．２ｋｂｐ以及２．５ｋｂｐ。其中仅２．２ｋｂｐ和２．５ｋｂｐ克隆与抗Ｐｃ９０单克隆抗体反应呈阳性。限制性内切酶酶切分析表明，２．２ｋｂｐ，２．５ｋｂｐ和１．６ｋｂｐ的克隆具有不同的特征。结论：２．２ｋｂｐ和２．５ｋｂｐ的克隆极有可能表达Ｐｃ９０蛋白。
OBJECTIVE: To construct a cDNA library of early trophozoites of Plasmodium cholerae and to screen positive clones expressing Pc90. Methods: GTC / CsCl method was used to extract RNA. The cDNA was cloned into λ-ZAP phage, screened with anti-Pc-HCPM serum three times, and then screened by anti-Pc90 monoclonal antibody. The positive clones were subjected to restriction endonuclease digestion Cut analysis. Results: The cDNA library has 1.9 × 106 recombinants. After screening with polyclonal antibodies against Pc90-HCPM, 16 positive clones were obtained with lengths of 0.6 kbp, 1.4 kbp, 1.6 kbp, 2.2 kbp and 2.5 kbp, respectively. Among them, only 2.2kbp and 2.5kbp clones reacted positively with anti-Pc90 monoclonal antibody. Restriction endonuclease analysis showed that 2.2kbp, 2.5kbp and 1.6kbp clones have different characteristics. Conclusion: The 2.2kbp and 2.5kbp clones are highly likely to express Pc90 protein.