五指山小型猪APEX1基因cDNA的克隆及生物信息学分析

cDNA Cloning and Bioinformatics Analysis of APEX1 Gene from Wuzhishan Miniature Pig

[目的]对五指山小型猪脱嘌呤嘧啶核酸内切酶cDNA进行克隆和生物信息学分析。[方法]以构建的五指山小型猪外周血白细胞cDNA文库为材料,采用菌落PCR方法,克隆得到APEX1基因全长cDNA,并运用生物信息学软件对该基因核苷酸序列及其编码蛋白的理化性质及二级结构等进行分析和预测。[结果]生物信息学分析表明,该cDNA全长1392bp,5′非翻译区长132bp,3′非翻译区长303bp,含有一个957bp的完整开放阅读框,编码318个氨基酸。该蛋白的分子量为35kD,等电点为8.05。比对分析表明,五指山小型猪APEX1基因的核酸序列及其氨基酸序列与人、小鼠、黑猩猩等哺乳动物具有较高的相似性。[结论]成功克隆了五指山小型猪A-PEX1基因cDNA,为进一步研究APEX1在动物体内的作用机制奠定了基础。

[ Objective] The study aimed to clone full-length cDNA of APEX1 gene from Wuzhishan miniature pig and bioinformaties analysis be carried out of this gene. [ Method ] Taking constructed cDNA library of the peripheral blood leucocytes from Wuzhishan miniature pig as material, the full-length cDNA of APEX1 gene was cloned by using colony PCR method. Nucleotide sequence of APEX1 gene,physicochemical properties and secondary structure of its encoding protein were analyzed and predicted by using bioinformatics software. [ Result] The bioinformatics analysis results showed that the full-length cDNA sequence was 1 392 bp, containing an open reading frame of 957 bp, flanked by 132 bp 5′UTR and 303 bp 3′UTR,encoded 318 amino acids. Relative molecular weight was 35 kD,pI was 8.05. Comparison analysis indicated that the nucleotide sequence and amino acid sequence of APEX1 gene were highly homologous to some mammals reported, such as human, mouse, himpauzee and other mammalian. [ Conclusion] The cDNA of APEX1 gene from Wuzhishan miniature pig was successfully cloned, it lays a foundation for further researching the action mechanism of APEX1 in the body of animal.