联系我们 加入收藏
 
年期检索 高级检索
33
2025年8月29日 星期五
农业生物技术学报  2021, Vol. 29 Issue (4): 753-763    DOI: 10.3969/j.issn.1674-7968.2021.04.013
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
虹鳟体色相关基因mitfa的克隆和表达分析
吴深基1, 黄进强1,*, 李永娟2, 张倩1, 潘玉财1, 王晓谰1
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
Cloning and Expression Analysis of Body Colour-related Gene mitfa in Rainbow Trout (Oncorhynchus mykiss)
WU Shen-Ji1, HUANG Jin-Qiang1,*, LI Yong-Juan2, ZHANG Qian1, PAN Yu-Cai1, WANG Xiao-Lan1
1 College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China;
2 College of Science, Gansu Agricultural University, Lanzhou 730070, China
全文: PDF (2974 KB)   HTML (1 KB) 
输出: BibTeX | EndNote (RIS)      
摘要 小眼畸形相关转录因子a基因(microphtalmia-associated transcription factor, mitfa)在动物体色形成过程中发挥重要的作用。为探讨mitfa基因在虹鳟(Oncorhynchus mykiss)体色变异中的调控作用,本研究采用cDNA末端快速扩增(rapid amplification of cDNA ends, RACE)技术首次获得了虹鳟mitfa基因的cDNA全长序列,并对其进行了生物信息学分析,同时利用qRT-PCR分析了野生型虹鳟(虹鳟)与黄色突变型虹鳟(金鳟)体色发生不同时期和不同组织中mitfa基因的表达差异。结果显示,虹鳟mitfa基因 (GenBank No. MT813117)的cDNA全长序列2 700 bp,其中开放阅读框1 212 bp,编码403个氨基酸,5'UTR 114 bp,3'UTR 1 374 bp;序列分析发现,虹鳟Mitfa蛋白序列与其他鱼类的同源性高于禽类、两栖类以及哺乳类,且基本螺旋-环-螺旋-亮氨酸拉链(basic helix-loop-helix-leucine zipper, bHLHZip)结构域在脊椎动物中高度保守;系统进化分析显示,虹鳟与褐鳟(Salmo trutta)的亲缘关系最近,与其他鱼类亲缘关系次之,与禽类、两栖类以及哺乳类亲缘关系最远;qRT-PCR分析表明,mitfa基因在虹鳟与金鳟胚胎及出膜后各时期均有不同程度的表达,均在受精期、4细胞期、16细胞期和桑葚期有较高的表达且显著高于其他各时期(P<0.05);虹鳟与金鳟相同时期的差异表达结果显示,该基因在受精期、16细胞期、桑葚期、囊胚期、体节期、心跳期、3 dph (days post hatch)、5 dph、7 dph、10 dph、1 M (month post hatch)和12 M背部皮肤中的表达差异极显著(P<0.01);mitfa基因在12月龄虹鳟和金鳟各组织中也均有表达,且都在背部皮肤中的表达量最高,在腹部的皮肤和肌肉、背部肌肉和眼中也有较高表达,其他组织该基因的表达相对较低。本研究结果表明,mitfa基因可能参与虹鳟色素细胞的形成过程,并与其体色变异密切相关。本研究结果为虹鳟体色变异分子调控机制研究及其体色遗传改良提供了基础资料。
服务
把本文推荐给朋友
加入我的书架
加入引用管理器
E-mail Alert
RSS
作者相关文章
吴深基
黄进强
李永娟
张倩
潘玉财
王晓谰
关键词 虹鳟体色小眼畸形相关转录因子a基因(mitfa)克隆表达分析    
Abstract:Microphtalmia-associated transcription factor a gene (mitfa) plays an important role in the formation of animal body colour. This study was conducted to explore the regulatory role of mitfa gene in the body colour variation between wild-type (WT) and yellow mutant (YM) rainbow trout (Oncorhynchus mykiss). The full-length sequence of mitfa gene of rainbow trout was obtained by rapid-amplification of cDNA ends (RACE) technique, and the bioinformatics analysis was performed. Furthermore, qRT-PCR was used to investigate the expression levels of mitfa in the different developmental stages and various tissues of WT and YM rainbow trout, respectively. The full-length cDNA of mitfa (GenBank No. MT813117) was 2 700 bp containing 114 bp 5'UTR, 1 374 bp 3'UTR, and 1 212 bp ORF encoding 403 amino acids. Sequence analysis showed that Mitfa protein sequence was more conservative in fishes than that in other vertebrates and the basic helix-loop-helix-leucine zipper (bHLHZip) structural domain was highly conserved among all vertebrates. These results were further confirmed by phylogenetic analysis. qRT-PCR analysis revealed that mitfa expressed at different levels during embryo and post hatch stages, and higher expression levels were detected at the fertilized-stage, 4-cell, 16-cell and multi-cell, which were significantly higher than that of other stages (P<0.05). Moreover, there were extremely significant differences (P<0.01) in the expression of mitfa gene at the same stages between WT and YM rainbow trout, e.g., at the fertilized-stage, 16-cell, multi-cell, blastula, somites, heartbeating, 3 dph (days post hatch), 5 dph, 7 dph, 10 dph, 1 M (month post hatch) and 12 M dorsal skin. mitfa gene expressed in all kinds of tissues of 12 M WT and YM rainbow trout, which expressed the highest amount in dorsal skin (P<0.05), it also highly expressed in ventral skin and muscle, dorasl muscle and eye, but relatively lower in other tissues. The above results indicated that the mitfa gene might be involved in the formation of pigment cells in rainbow trout and closely related to the body colour variation. The results of this study provide basic data for the molecular regulation mechanism of body colour variation and the genetic improvement of body colour in rainbow trout.
Key wordsRainbow trout    Body colour    Microphtalmia associated transcription factor a gene (mitfa)    Cloning    Expression analysis
收稿日期: 2020-08-13      出版日期: 2021-04-01
ZTFLH:  S917.4  
基金资助:国家自然科学基金(31760755)
通讯作者: * huangjq@gsau.edu.cn   
引用本文:   
吴深基, 黄进强, 李永娟, 张倩, 潘玉财, 王晓谰. 虹鳟体色相关基因mitfa的克隆和表达分析[J]. 农业生物技术学报, 2021, 29(4): 753-763.
WU Shen-Ji, HUANG Jin-Qiang, LI Yong-Juan, ZHANG Qian, PAN Yu-Cai, WANG Xiao-Lan. Cloning and Expression Analysis of Body Colour-related Gene mitfa in Rainbow Trout (Oncorhynchus mykiss). 农业生物技术学报, 2021, 29(4): 753-763.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/10.3969/j.issn.1674-7968.2021.04.013     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2021/V29/I4/753
 
版权所有 © 2014 《农业生物技术学报》编辑部   京ICP备11035905号-3
地址:北京市海淀区圆明园西路2号中国农业大学生命科学楼1053室 邮编:100193
电话:010-62733684 传真:010-62731615 E-mail: nsjxb@cau.edu.cn