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目的 :阴道毛滴虫黏附蛋白AP33基因的克隆与序列分析。方法 :提取阴道毛滴虫mRNA ,逆转录合成cDNA ,PCR得到阳性条带 ,将其克隆到pUCm T载体进行PCR、酶切及测序分析 ,并与GenBank中核苷酸序列进行同源性分析。结果 :阳性克隆经酶切鉴定 ,目的基因片段长度为 92 7bp。测序结果目的基因与阴道毛滴虫黏附蛋白AP33 1、阴道毛滴虫黏附蛋白AP33 3、阴道毛滴虫黏附蛋白AP33 2分别具 99%、97%和 94 %的同源性。结论 :成功克隆出阴道毛滴虫AP33基因序列 ,与已公布的AP33序列有很高的同源性。这为今后该蛋白的原核表达和大量制备提供一定的基础。
Objective: Cloning and sequence analysis of Trichomonas vaginalis adhesion protein AP33 gene. Methods: Trichomonas vaginalis mRNA was extracted and cDNA was reverse transcribed. The positive clones were obtained by PCR. The positive clones were cloned into pUCm T vector and analyzed by PCR, restriction analysis and sequencing. The homology was also analyzed with GenBank. Results: The positive clones were identified by restriction enzyme digestion and the length of the target gene fragment was 92 7 bp. The sequencing results showed that the target gene shared 99%, 97% and 94% homology with Trichomonas vaginalis adhesion protein AP33 1, Trichomonas vaginalis adhesion protein AP33 3 and Trichomonas vaginalis adhesion protein AP33 2, respectively. Conclusion: The sequence of AP33 gene of Trichomonas vaginalis was successfully cloned, which has high homology with published AP33 sequence. This provides a certain basis for the prokaryotic expression and mass preparation of the protein in the future.