浅色黄姑鱼Ⅰ型胶原蛋白α2(COL1A2)基因的克隆及表达分析  

Cloning and mRNA Expression Analysis of TypeⅠCollagen Alpha 2 Gene from Chu’s Croaker(Nibea coibor)

作　　者：荣华 凌钰 豆腾飞 黄英[2] 王晓雯[2] 孔令富[2] 毕保良[2] 王金浩 RONG Hua;LING Yu;DOU Tengfei;HUANG Ying;WANG Xiaowen;KONG Lingfu;BI Baoliang;WANG Jinhao(College of Agriculture,Xiangyang Polytechnic,Xiangyang 441050,China;Faculty of Animal Science and Technology,Yunnan Agricultural University,Kunming 650201,China;Yunnan Huaheng Testing Technology Co.,Ltd.,Kunming 650201,China)

机构地区：[1]襄阳职业技术学院农学院,湖北襄阳441050 [2]云南农业大学动物科学技术学院,云南昆明650201 [3]云南华衡检测技术有限公司,云南昆明650201

出　　处：《云南农业大学学报（自然科学版）》2023年第5期786-794,共9页Journal of Yunnan Agricultural University:Natural Science

基　　金：云南省基础研究计划面上项目(202201AT070251);云南省重大科技专项计划项目(202202AE090018);广东省基础与应用基础研究基金项目(2021A1515110179);湖北省自然科学基金一般面上项目(2023AFB1016)。

摘　　要：【目的】通过克隆浅色黄姑鱼(Nibea coibor)Ⅰ型胶原蛋白α2(COL1A2)基因,分析其分子特征、组织分布及表达差异,为鱼类胶原蛋白代谢机制研究奠定理论基础。【方法】通过cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)技术获得浅色黄姑鱼COL1A2基因全长序列,利用生物信息学分析方法对其进行系统分析,探讨其分子特征及组织表达谱。【结果】COL1A2 cDNA序列长5826 bp,编码1352个氨基酸。BLAST同源性比对和系统进化树分析表明:浅色黄姑鱼与大黄鱼(Larimichthys crocea)同属石首鱼科(Sciaenidae),亲缘关系最近,且两者COL1A2的氨基酸序列同源性最高(96.60%)。生物信息学分析显示:COL1A2多肽链包含脊椎动物Ⅰ型胶原蛋白的所有特征,如相同的信号肽、C-前肽和N-前肽结构域以及三重螺旋结构域等。qRT-PCR结果显示:鱼鳔中COL1A2基因的表达显著高于其他组织(P<0.05),且胶原蛋白在不同组织中的分布规律与其一致。【结论】本研究首次克隆了浅色黄姑鱼COL1A2基因(GenBank登录号:MK641513),该基因的组织表达特异性是引起胶原蛋白差异化沉积的原因之一。研究结果有助于深入探究胶原蛋白的生物合成途径及调控机制。[Purpose]This study cloned cDNA of COL1A2 gene from Niber coibor,analyzed the molecular characteristics,tissue distribution and expression of collagen,to lay a theoretical foundation for the study of collagen metabolism mechanism in fish.[Methods]The full-length sequence of COL1A2 gene of N.coibor was obtained through rapid amplification of cDNA ends(RACE)technology,and then systematically analyzed using bioinformatics analysis method to explore its molecular characteristics and tissue expression.[Results]The COL1A2 cDNA was 5826 bp in full-length and encoded a polypeptide of 1352 amino acids.BLAST homology comparison and phylogenetic tree analysis revealed that the N.coibor and Larimichthys crocea should belong to Sciaenidae,having the closest relatives,and the amino acid sequence homology of COL1A2 was the highest(96.60%).Bioinformatics analysis showed that COL1A2 contained all the characteristics of type I collagen in vertebrates identified,such as the same signal peptides,C-propeptide and N-propeptide domains,triple helix domains and so on.qRT-PCR results showed that the expression level of COL1A2 in the swim bladder of N.coibor was significantly higher than that in other tissues(P<0.05).It was consistent with the distribution pattern of collagen in different tissues.[Conclusion]The COL1A2 gene of N.coibor(GenBank accession number:MK641513)is cloned for the first time.The tissue expression specificity of this gene is one of the reasons for the differential depositon of collagen.The results of the study will help to further study the collagen biosynthetic pathway and regulation mechanism in N.coibor.

关 键 词：浅色黄姑鱼 COL1A2 CDNA克隆 表达分析 组织分布 

分 类 号：Q959.483[生物学—动物学]