锦鲤TYR基因克隆、序列分析及在不同组织中表达  

Molecular Cloning,Sequence Analysis and Expression Detection of TYR from Koi Carp

作　　者：熊钢[1] 王宇[1] 王晓清[2] 刘臻[3] 马晓[2] 张建国[1] 卿爱东[1] 

机构地区：[1]湖南生物机电职业技术学院动科系,湖南长沙410127 [2]湖南农业大学动物科技学院,湖南长沙410128 [3]长沙学院生物工程与环境科学系,湖南长沙410003

出　　处：《生物技术》2013年第3期8-12,共5页Biotechnology

基　　金：湖南省教育厅科学研究项目("锦鲤色素细胞发育和酪氨酸酶基因研究";11C0792)资助;高级研究员王宇的项目资金资助~~

摘　　要：目的:克隆锦鲤TYR cDNA基因,分析基因生物信息及几中组织中表达情况。方法:采用RT-PCR、基因3’和5’扩增技术,从锦鲤体表组织克隆TYR cDNA全长基因,用生物软件对基因序列进行基因生物信息分析;采用RT-PCR方法检测TYR基因在锦鲤几种组织的表达情况。结果:成功克隆锦鲤TYR cDNA基因,基因长1 999bp,包含26 bp的5’UCR序列,371 bp的3’UCR序列,1 602 bp开放阅读框,编码533个氨基酸,蛋白质分子量60.426kDa,预测TYR蛋白有6个α结构;对其构建的基因系统进化树与传统形态分类相吻合;并在锦鲤皮肤、肌肉、组织、鳍和眼组织均检测到基因的表达。结论:该研究为进一步探讨锦鲤TYR基因、TYR蛋白功能和黑色素的形成变化机理奠定基础。Objective： It was to clone and analyse the TYR eDNA gene of Koi grap, detect the gene expression in several tissue. Method： By using RT - PCR method, TYR gene of Koi carp was cloned from surface tissues＇ s totoal RNA. C, ene sequences was analysed with bio- logical software, and detected in several organizations expression with RT - PCIL Result ：Sequencing results showed that the gene included 1 999 bp. Nucleotide sequence analysis showed that TYR cDNA includes a 26bp long 5＇ - untranslated region（ UTR）, ORF encoding 533 amino acids, and a 371bp long 3＇ URT ; the molecular weight predicted is 60. 426 kDa; TYR has six a tertiary structure; The evolutiontree built by kimura way from the TYR cDNA homology of Koi carp and other animals was consistent with that of traditional taxonomic re- suits ; The TYR mRNA was detected in skin, muscle, fins and eye tissue of Koi carp. （~Ola^lusion： The research laid a foundation for fur- ther study on the metabolism of Koi carp TYR gene, the protein function and The mechanism of melanin formation and change.

关 键 词：锦鲤 TYR 基因克隆 生物息信分析 组织表达 

分 类 号：Q785[生物学—分子生物学] Q786