摘要
利用PCR和RACE方法首次克隆了编码草鱼肌肉Ⅰ型胶原蛋白的α1基因(COL1A1)的cDNA全长序列,为5772bp,其开放阅读框为4347bp,编码1448个氨基酸。BLAST同源性分析结果显示,草鱼COL1A1基因的氨基酸序列与斑马鱼、金鱼同源性较高,分别为93.90%和93.60%,呈现出较高的保守性。系统进化树分析表明,该基因与斑马鱼、金鱼处于同一支,亲缘性最近。生物信息学分析显示,草鱼COL1A1蛋白质的相对分子质量为137.2ku,理论等电点是5.44,为α螺旋β折叠三重螺旋结构蛋白。有18段三重螺旋重复域,22段低复杂度域,17个功能域。COL1A1蛋白有33～69,1249～1355两个绑定钙的区域和62～104一个绑定锌的区域。利用半定量RT-PCR方法检测的组织表达结果表明,COL1A1基因在草鱼肌肉、肠道、肝胰脏、鳃、皮肤、鳍条、肾脏和脾脏8个组织中均有表达,其中在皮肤、鳃、肾脏、鳍条组织中的mRNA表达量高于其他4个组织(P<0.05)。
TypeⅠcollagen, as a number of collagen family, is the most abundant collagen and major components of the extracellular matrices of all metazoan life, and plays crucial roles in differentiation, formation of collagen fibers and tissue remodeling after injury, etc. Type Ⅰ collagen alpha 1（COL1A1） cDNA of grass carp （Ctenopharyngodon idella） was isolated through the RT-PCR and RACE approaches. The cDNA was 5 772-bp in length, including a 4 347-bp CDS （coding sequence） and encoded a polypeptide of 1 449 aa. The homology of COL1A1 amino acid with relative species （zebrafish, goldfish, etc.） was as high as 93% with zebra fish and goldfish. The protein peptide molecular weight was 137.2 ku and theoretical pI was 5.44 using ProtParam software on line. The protein peptides of COL1A1 possessed 6 α-helixes, 12 β-sheets, others of ruleless coil regions, and 18 regions of triple helical repeats, 22 low complexity regions, 17 function domains. There were two calcium-binding sites and one zinc-binding site in the COL1A1 protein peptide. COL1A1 mRNA was determined in all the tested 8 tissues （muscle, intestine, hepatopancreas, gill, skin, fin, kidney and spleen） of grass carp by semi- quantitative RT-PCR, and the mRNAs expression in gill, kidney, skin and fin significantly higher than other tissues （P0.05）. The structure and bioinformatics characteristics of the COL1A1 from grass carp may help to further understand the function of COL1A1 gene in the repair process of damaged tissue in the grass carp.
出处
《水产学报》
CAS
CSCD
北大核心
2012年第6期849-858,共10页
Journal of Fisheries of China
基金
广东省自然科学基金自由申请项目(10151038001000004)
现代农业产业体系建设专项项目(nycytx-49)
广东省海洋渔业科技推广项目(A200901D04)
