虹鳟Scarb1基因克隆、生物信息学及组织表达分析  

Cloning,Bioinformatics and Tissues Expression Analysis of Scarb1 Gene in Rainbow Trout

作　　者：张东强 黄进强[1] 李永娟[1,2] 吴深基 赵璐 宋玉芳 ZHANG Dongqiang;HUANG Jinqiang;LI Yongjuan;WU Shenji;ZHAO Lu;SONG Yufang(College of Animal Science and Technology,Gansu Agricultural University,Lanzhou 730070,China;College of Science,Gansu Agricultural University,Lanzhou 730070,China)

机构地区：[1]甘肃农业大学动物科学技术学院,兰州730070 [2]甘肃农业大学理学院,兰州730070

出　　处：《西北农业学报》2024年第6期1008-1018,共11页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(31760755);甘肃农业大学学科团队项目(GAU-XKTD-2022-23)。

摘　　要：清道夫受体B类成员1(scavenger receptor class B member 1,Scarb1)作为细胞表面的膜受体蛋白,在动物体色形成过程中发挥重要作用。为了解Scarb1基因在虹鳟(Oncorhynchus mykiss)体色形成中的作用,通过RACE技术克隆虹鳟Scarb1基因的cDNA全长,并运用生物信息学方法分析该基因及其序列结构特征,同时使用实时定量PCR(qRT-PCR)检测Scarb1基因在虹鳟、金鳟及其杂交F_(1)代不同发育阶段和不同组织中的表达情况。结果显示,Scarb1基因cDNA序列全长为2032 bp,开放阅读框1479 bp,编码492个氨基酸,预测分子质量为55.59 ku,且存在保守的CD36结构域和2个跨膜区。序列同源性分析显示,虹鳟与其他硬骨鱼类的氨基酸序列相似度为71.69%~98.58%;进化分析发现虹鳟与大马哈鱼亲缘关系最近,与哺乳动物和两栖动物亲缘关系最远。qRT-PCR检测结果表明,在虹鳟与金鳟胚胎期及出膜后各发育阶段中Scarb1基因均有不同程度表达,且表现为受精期至桑葚期的表达显著高于其他时期(P<0.05),对虹鳟与金鳟同一时期的差异分析发现该基因在胚胎期及7 dph(days post hatch)、1 M(month post hatch)、2 M和3 M时期中表达存在显著差异(P<0.01)。Scarb1基因在虹鳟与金鳟背部皮肤和背部肌肉等色素沉着性组织中表达量较高,其中在金鳟背部皮肤的表达量显著高于虹鳟(P<0.01)。此外,Scarb1基因在杂交F_(1)代不同发育时期中的表达规律与双亲一致;在不同组织中,该基因在杂交F_(1)代背部皮肤中的表达量介于双亲之间。研究结果表明,Scarb1基因与虹鳟体色形成有着密切关系,且可能在金鳟黄色体色形成过程中发挥重要作用。Scavenger receptor class B member 1(Scarb1),a cell surface membrane receptor protein,plays an important role in the formation of skin color in animals.To study the function of Scarb1 gene in the formation of skin color of rainbow trout(Oncorhynchus mykiss),we obtained the full-length cDNA sequence of rainbow trout Scarb1 gene by RACE technique,and analyzed its sequence features by bioinformatics.The quantitative real-time PCR(qRT-PCR)was used to detect the expression of Scarb1 gene at different developmental stages and tissues of wild-type rainbow trout(WT),yellow mutant rainbow trout(YM)and their hybrid F_(1) generation.The results showed that the full length of Scarb1 cDNA was 2032 bp,with an open reading frame of 1479 bp,encoding 492-amino acid protein.The molecular mass of the protein was approximately 55.59 ku and conserved CD36 structural domain and two transmembrane regions.The homology analysis showed that the amino acid sequence similarity of Scarb1 between rainbow trout and other bony fishes was 71.69%to 98.58%.The phylogenetic analysis showed that rainbow trout had the closest relationship with salmon,and the farthest relationship with mammals and amphibians.The qRT-PCR analysis showed that Scarb1 gene was expressed at embryo and post hatch stages of WT and YM,and the expression at fertilization stage,4-cell,16-cell and multi-cell was significantly higher than that at other stages(P<0.05).Additionally,the expression of Scarb1 gene was extremely significant differences(P<0.01)at the embryonic stage,7 dph(days post hatch),1 M(month post hatch),2 M and 3 M stage of WT and YM.The expression of Scarb1 gene was higher at the dorsal skin and dorsal muscle,and the dorsal skin of YM was significantly higher than that of WT(P<0.01).Moreover,the expression pattern of Scarb1 gene at different developmental stages of F_(1) generation was consistent with that of both parents,and the expression of the gene in the dorsal skin of the F_(1) generation was intermediate between the both parents.All these results suggest t

关 键 词：虹鳟 体色 清道夫受体B类成员1(Scarb1) 基因克隆 表达分析 

分 类 号：S917.4[农业科学—水产科学]