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目的 :获取阴道毛滴虫硫氧还原蛋白过氧化物酶的cDNA克隆 ,研究其还原烷基氢过氧化物和氢过氧化物的抗氧化作用 ,以及调节由氢过氧化物介导的信号转换。 方法 :提取阴道毛滴虫的总RNA ,用λTripIEx2噬菌体载体构建cDNA表达文库 ,阳性质粒cDNA克隆进行测序 ,并用生物软件RPS Blast,NCBIBlast和ClustalW等对 6个读码框架进行序列分析。 结果 :获得一株开放阅读框为 5 88对碱基的cDNA克隆 ,推测肽链有 196个氨基酸。序列分析表明 ,该肽链与衣滴虫过氧化物氧化还原酶 (Prx1蛋白 )及人的自然杀伤增强因子B分别有 5 6 %和 6 1%的同源性 ;除了两个高度保守的半胱氨酸作用基序外 ,还有蛋白激酶C磷酸化和酪氨酸激酶磷酸化作用基序 ,N 豆蔻酰化作用位点 ,脂质运载蛋白信号位点等。 结论 :该蛋白有 >5 6 %的可能性位于胞浆 ,和典型 2 CysPrx亚家族有很高 (5 6 %～ 6 2 % )的同源性 ,很可能是其中的一个新成员。
OBJECTIVE: To obtain the cDNA clone of Trichomonas vaginalis thioredoxin peroxidase and to study its antioxidative effect on reduction of alkyl hydroperoxides and hydroperoxides, as well as on the regulation of signal transduction mediated by hydroperoxides . Methods: The total RNA of Trichomonas vaginalis was extracted, cDNA expression library was constructed with λTripIEx2 phage vector, and the positive plasmid cDNA clone was sequenced. Sequence analysis was performed on six reading frames using biological softwares RPS Blast, NCBIBlast and ClustalW. RESULTS: A cDNA clone of 5 88 bp was obtained with an open reading frame of 196 amino acids. Sequence analysis showed that the peptide chain shared 56% and 61% homology with Trichomonas peroxidase oxidoreductase (Prx1) and human natural killer enhancer B, respectively; except for two highly conserved half Cystine action motif, but also protein kinase C phosphorylation and tyrosine kinase phosphorylation motifs, N myristoylation sites, lipid signaling sites and other proteins. CONCLUSION: This protein is located in the cytoplasm with a> 56% chance of being highly homologous to the typical 2 CysPrx subfamily and is likely to be a new member of the 2 CysPrx subfamily.