虹鳟酪氨酸酶基因的生物信息学分析及其在不同发育阶段和组织中的表达分析

doi:10.16431/j.cnki.1671-7236.2021.05.002

摘要/Abstract

摘要： 酪氨酸酶（tyrosinase，Tyr）基因在调控动物色素合成的过程中发挥关键作用。为了解tyr基因与虹鳟（Oncorhynchus mykiss）体色变异的关系，本研究对虹鳟tyr-1和tyr-2基因进行了蛋白序列分析，并利用实时荧光定量PCR检测两种基因在野生型虹鳟（虹鳟）和黄色突变型虹鳟（金鳟）体色发生的19个不同时期和成鱼13种不同组织中的表达差异。蛋白序列分析结果表明，Tyr-1和Tyr-2都是亲水性蛋白，且蛋白结构主要是无规则卷曲和α-螺旋；实时荧光定量PCR结果显示，tyr-1基因在胚胎各时期均有表达，tyr-1基因表达量在虹鳟受精期最高，桑椹期次之，且显著高于其他时期（P<0.05）；tyr-1基因表达量在金鳟16-细胞期最高，且显著高于其他时期（P<0.05）；tyr-2基因分别从虹鳟囊胚期和金鳟原肠期开始表达，表达量均在心跳期达到最高且显著高于其他时期（P<0.05）。出膜后，tyr-1、tyr-2基因分别在虹鳟5日龄和金鳟3月龄表达量最高。在胚胎各时期中，tyr-1、tyr-2基因在虹鳟中表达量普遍高于金鳟，出膜后普遍低于金鳟。在各组织中，tyr-1基因在皮肤、眼睛、肾脏、肝脏等组织中有较高表达，tyr-2基因主要在皮肤、眼睛中有较高表达，在其他组织中表达量较低。以上结果表明，tyr-1和tyr-2基因与虹鳟体色变异具有一定的相关性，为深入阐明虹鳟体色变异机制和体色遗传改良奠定了理论基础。

关键词: 虹鳟; 体色; 酪氨酸酶基因; 生物信息学分析; 表达分析

Abstract: Tyrosinase (Tyr) gene plays a key role in the process of regulating animal pigment synthesis.In order to understand the relationship between tyr gene and body colour variation of rainbow trout,in this study,the protein sequence analysis of tyr-1 and tyr-2 genes in rainbow trout was performed,the expression of 19 different development stages and 13 different tissues was determined by Real-time quantitative PCR in wild-type rainbow trout (WT) and yellow mutant rainbow trout (YT).Protein sequence analysis results showed that Tyr-1 and Tyr-2 proteins were both hydrophilic proteins and the structure was mainly random coil and alpha helix;Real-time quantitative PCR results showed that tyr-1 gene was expressed in WT and YT embryos at all stages.In WT,the expression of tyr-1 gene was the highest in the fertilized-stage,followed by the multi-cell stage,which was significantly higher than other stages (P<0.05),In YT,the expression of tyr-1 gene expression was the highest at the 16-cell,which significantly higher than other periods (P<0.05);tyr-2 gene was expressed from the WT blastula stage and the YT gastrula stage,the expression level reached the highest in the heartbeating stage,which was significantly higher than other periods (P<0.05).The expression levels of tyr-1 and tyr-2 genes were the highest in WT at 5 days post hatch and YT at 3 months post hatch.The expression levels of tyr-1 and tyr-2 genes in WT were higher than those in YT at embryonic stage and generally lower than those in YT during post hatch.In adult fish tissues,tyr-1 gene was highly expressed in skin,eyes,kidney,liver and other tissues,the tyr-2 gene was highly expressed in skin and eyes,and it was rarely expressed in other tissues.The above results indicated that tyr-1 and tyr-2 genes had a certain correlation with the body color variation of rainbow trout,which provided a theoretical basis for in-depth elucidation of the body color variation mechanism and body color genetic improvement of rainbow trout.

Key words: rainbow trout; body colour; tyr gene; bioinformatics analysis; expression analysis

中图分类号:

Q786

王晓谰, 吴深基, 黄进强, 李永娟, 潘玉财, 张倩. 虹鳟酪氨酸酶基因的生物信息学分析及其在不同发育阶段和组织中的表达分析[J]. 中国畜牧兽医, 2021, 48(5): 1516-1524.

WANG Xiaolan, WU Shenji, HUANG Jinqiang, LI Yongjuan, PAN Yucai, ZHANG Qian. Bioinformatics and Expression Analysis of Tyrosinase Gene at the Different Stages and Tissues of Rainbow Trout (Oncorhynchus mykiss)[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(5): 1516-1524.

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