鹿茸X型胶原基因的全长cDNA克隆及序列分析

Molecular cloning and expression analysis of antler collagen X

采用RT-PCR和RACE技术,从鹿茸软骨细胞总RNA中扩增克隆了梅花鹿X型胶原(collagen X,colX)全长cDNA序列,并进行了序列特性分析。序列分析结果表明,梅花鹿colX全长cDNA序列为3135nt,其中5′非编码区96nt、3′非编码区1014nt和2022nt的开放阅读框编码674个氨基酸的鹿colX前体蛋白,与牛、猪、犬、人类和小鼠等脊椎动物colX氨基酸序列之间的同源性超过82%。按该基因氨基酸序列构建了进化树,揭示其与牛基因更加同源。

collagen X （col X） plays an important role in the enchondral bone formation. The methods of isolation and culture sika antler chondrocytes were established. Sika antler chondrocytes were cultured. Full-length cDNA of sika col X gene was amplified and cloned from total RNA of sika antler chondrocytes by the RT-PCR and RACE methods. The cDNA sequence was 3 135 nt in length, including 96 nt of 5＇untranslated region, 2 022 nt of coding region and 1 014 nt of 3＇untranslated region. The coding region encoded a protein precursor with 674 amino acid residues. Amino acid identities of col X between Sika and other vertebrates including cow, pig, dog, human and mouse were more than 82%. Phylogenetic tree analysis based on the amino acid sequences showed that col X of sika and cow was within one clade.