阴道毛滴虫氢化酶体腺苷酸激酶基因的克隆及序列分析

目的-克隆阴道毛滴虫氢化酶体腺苷酸激酶(AK)基因，并测定其序列，进行序列分析。方法-根据AK基因已知序列设计合成一对引物，应用PCR技术从阴道毛滴虫基因组DNA中扩增出AK基因，并将其克隆入pMD18-T simple载体。阳性克隆的重组质粒经酶切及PCR鉴定后，用双脱氧链末端终止法进行基因序列测定。应用BLAST软件辅助分析所测基因与Genbank中阴道毛滴虫氢化酶体AK序列的同源性。结果-PCR扩增得到特异的阴道毛滴虫氢化酶体腺苷酸激酶基因序列。酶切及PCR鉴定获得了正确的PT-AK重组质粒。测序表明，所克隆的AK基因大小为690bp，编码229个氨基酸。序列分析表明，所测基因与Genbank中阴道毛滴虫氢化酶体AK序列具有高度同源性(99．9％)。结论-克隆了阴道毛滴虫氢化酶体腺苷酸激酶基因，序列测定及同源性分析表明，所测基因与Genbank中阴道毛滴虫氢化酶体AK序列具有高度同源性。

Cloning and Sequence Analysis of Hydrogenosomal Adenylate Kinase (AK) of Trichomonas vaginalis

Objective To clone the gene of AK and analyse encoding sequence of AK gene of Trichomonas vaginalis. Methods A pair of primers was designed according to the known sequence of AK gene. The AK gene fragment was amplified by PCR, and ligated to a sequencing, pMD18-T simple vector. Positive recombinants were detected by PCR, digested by restriction enzyme and sequenced with dideoxy nucleotide chain termination method. Result The sequence analysis showed that the sequenced protein showed 99.9% of homology with the AK amino acid sequence in Genbank. Conclusion The gene of AK has high homology with the AK gene sequence in Genbank.