猪TRIM38基因的克隆与序列分析

Cloning and sequence analysis of the porcine TRIM38 gene

为了克隆三基序蛋白38(tripartite motif 38,TRIM38)基因,并对该基因编码蛋白进行序列分析及生物信息学功能预测,试验采用PCR方法扩增猪TRIM38基因,并将其连接至pMD-18T载体上;采用多种在线软件预测猪TRIM38蛋白的理化性质和生物信息学功能。结果表明:猪TRIM38基因序列长度为1398 bp。大白猪TRIM38基因的核苷酸序列与欧亚野猪的相似性为100%,亲缘关系最近,为同一物种;与非洲跳鼠的相似性为70.1%,亲缘关系最远。猪TRIM38基因编码的蛋白质由465个氨基酸分子组成,分子量为32.27 ku,理论等电点(pI)为6.18,呈酸性。平均亲水系数(GRAVY)为-0.502,不稳定系数为60.89,是一种不稳定的亲水性蛋白质;猪TRIM38蛋白具有26个磷酸化位点,并含有16个糖基化位点。大白猪TRIM38蛋白主要存在于细胞核中;该蛋白无信号肽,不属于分泌蛋白且为膜外蛋白;猪TRIM38蛋白二级结构主要以α-螺旋为主,三级结构与二级结构相符。说明TRIM38基因可以调控PRRSV的复制。

In order to clone the tripartite motif 38(TRIM38)gene and perform sequence analysis and bioinformatics function prediction of the protein encoded by this gene,the porcine TRIM38 gene was amplified by PCR and connected to the pMD-18T vector.The physicochemical properties and bioinformatics functions of pig TRIM38 protein have been predicted through various online software.The results showed that the sequence length of porcine TRIM38 gene was 1,398 bp;the sequence of large white porcine TRIM38 gene was 100%similar to Eurasian wild boar,which was the closest relative and was the same species,and the similarity with African jumping mouse was 70.1%,which was the farthest relative.The protein encoded by porcine TRIM38 gene is composed of 465 amino acid molecules with molecular weight 32.27 ku and a theoretical isoelectric point(PI)of 6.8,indicating acidity.The average hydrophilicity coefficient(CRAVY)of this protein was-0.502,and the instability coefficient was 60.89,indicating that it was an unstable hydrophilic protein.The porcine TRIM38 protein had 26 phosphorylation sites and contained 16 glycosylation sites.The porcine TRIM38 protein mainly existed in the nucleus;the protein had no signal peptide,is not a secreted protein and was an extracellular protein.The secondary structure of porcine TRIM38 protein was mainly dominated byα-helices,and the tertiary structure model was consistent with the results of the secondary structure.The results indicated that the TRIM38 gene could regulate thereplicationof PRRSV.