猪泛素C端水解酶L1基因的克隆与序列分析

从人泛素C端水解酶L1(UCH_L1)基因出发,在dbEST数据库中同源搜索,找到1条与人UCH_L1基因编码氨基酸同源性较高且在香猪背最长肌中表达的EST(BM194679).通过电子克隆和进一步RT-PCR试验验证,获得猪UCH_L1基因全长cDNA序列,其全长 1 105 bp,开放阅读框(ORF)位于60～728 bp,编码223个氨基酸.同源性分析结果表明,与人、鼠UCH_L1基因cDNA编码区(CDS)同源性分别为91.2%和86.5%,蛋白质序列同源性均为96.6%.对该基因编码蛋白的结构和功能预测显示含有2个典型的疏水性区域,不含有信号肽,存在1个UCH_L1(pfam01088)保守结构域和多个磷酸化位点,属UCH_L1蛋白家族,故将该基因命名为猪泛素C端水解酶L1基因(登录号AY495532).

Cloning and Sequence Analysis of Ubiquitin C-Terminal Hydrolase L1 Gene of Pig

The human ubiquitin C-terminal hydrolase L1(UCH_L1) gene cDNA was used to search the dbEST database for homologous porcine expressed sequence tags(ESTs).A matching EST(BM194679) previous found to express Xiang pig longissimus dorsi muscle was identified.Through in silico cloning and RT-PCR with total RNA from Landrace pig longissimus dorsi muscle as the template,a(1 105 bp) cDNA fragment was amplified and subsequently sequenced.Sequence analysis revealed that this(1 105 bp) fragment contained a(669 bp) open reading frame and shared the highest homogenecitis at 91.2% and 86.5% with human and mouse homologues,respectively.The deduced peptide of 223 amino acid residues showed 96.6% homogenety to the orthologous of human and mouse,and contained a classical UCH_L1(pfam01088)domain,2 typical transmembrane domains and 3 biological-meaning sites,but with no signal peptide.Based on these characteristics this gene was nominated as pig UCH_L1 gene(GenBank(acession) number AY495532).