| 红鳍笛鲷视杆蛋白和长波敏感视蛋白的基因克隆及表达规律分析 |
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| 引用本文： | 陈子钊,刘雪媚,许振民,梁秋璐,王中铎,郭昱嵩.红鳍笛鲷视杆蛋白和长波敏感视蛋白的基因克隆及表达规律分析[J].南方农业学报,2022,53(11):3285-3296. |
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| 作者姓名： | 陈子钊 刘雪媚 许振民 梁秋璐 王中铎 郭昱嵩 |
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| 作者单位： | 1. 广东海洋大学水产学院/南海水产经济动物增养殖广东普通高校重点实验室, 广东湛江 524088;2. 广东省水产动物病害防控与健康养殖重点实验室, 广东湛江 524088 |
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| 基金项目： | 广东海洋大学创新强校项目(230419055)国家自然科学基金项目(31972794) |
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| 摘 要： | 【目的】克隆红鳍笛鲷(Lutjanus erythropterus)视杆蛋白(RH1)基因和长波敏感视蛋白(LWS)基因,并分析其在早期发育不同阶段和成鱼不同组织的表达规律,为探究笛鲷属鱼类适应从表层浮游逐步转为下层底栖光环境生活的过程提供理论基础。【方法】利用RACE克隆红鳍笛鲷RH1基因和LWS基因cDNA全长,利用生物信息学软件对2个基因序列及编码的氨基酸序列进行分析,并利用实时荧光定量PCR检测其在红鳍笛鲷6个胚胎发育时期(原肠下包1/2期、胚孔封闭期、视囊期、晶体出现期、心脏跳动期、孵化出膜期)和5个仔鱼发育时期(1、3、10、15和20 d),以及成鱼不同组织(脑脏、心脏、肝脏、肌肉、黑色皮肤、红色皮肤、胃和视网膜)的表达规律。【结果】红鳍笛鲷RH1基因和LWS基因cDNA全长分别为1723 bp和1302 bp,其中RH1基因具有465 bp的3'-UTR和196 bp的5'-UTR,开放阅读框(ORF)长度为1062 bp,编码353个氨基酸残基;LWS基因具有213 bp的3'-UTR和15 bp的5'-UTR,ORF长度为1074 bp,编码357个氨基酸残基。系统发育分析结果显示,2个视蛋白基因均先与鲈形目等鱼类聚为一支,再与鳉形目、鲽形目和鲤形目等聚为一支,最后再与陆生脊椎动物聚为一支。实时荧光定量PCR检测结果显示,RH1基因在孵化出膜后15和20 d的相对表达量显著高于其他时期(P<0.05,下同),LWS基因在孵化出膜后20 d的相对表达量显著高于其他时期;2个视蛋白基因在成鱼不同组织的表达模式相似,在视网膜上的相对表达量均显著高于其他组织。【结论】红鳍笛鲷RH1和LWS基因的表达具有组织特异性,且在早期发育阶段的表达水平与红鳍笛鲷的生活习性变化相关,表明红鳍笛鲷RH1和LWS基因表达与其生活光环境的变化密切呼应,在早期发育变态阶段的光线调节,以及适应黑暗环境等方面发挥重要作用。
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| 关 键 词： | 红鳍笛鲷 RH1基因 LWS基因 基因克隆 表达分析 |
| 收稿时间： | 2022-05-31 |
Cloning and expression regulation of rhodopsin and long-wave sensitive opsin in Lutjanus erythropterus |
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| CHEN Zi-zhao,LIU Xue-mei,XU Zhen-min,LIANG Qiu-lu,WANG Zhong-duo,GUO Yu-song.Cloning and expression regulation of rhodopsin and long-wave sensitive opsin in Lutjanus erythropterus[J].Journal of Southern Agriculture,2022,53(11):3285-3296. |
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| Authors: | CHEN Zi-zhao LIU Xue-mei XU Zhen-min LIANG Qiu-lu WANG Zhong-duo GUO Yu-song |
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| Affiliation: | 1. Fisheries College, Guangdong Ocean University/Key Laboratory of Aquaculture in South China Sea for Aquatic Economic Animal of Guangdong Higher Education Institutes, Zhanjiang, Guangdong 524088, China;2. Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, Zhanjiang, Guangdong 524088, China |
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| Abstract: | 【Objective】In order to provide a theoretical basis for exploring the process of snapper adaptation from surface plankton to bottom benthic light environment. Therhodopsin(RH1) gene and long-wave sensitive opsin(LWS) gene of Lutjanus erythropterus were cloned and their expression law were analyzed in different stages of early development and in different tissues of adult fish.【Method】The full-length cDNA of RH1 gene and LWS gene of L. erythropterus were cloned by RACE technology. Thesequence of the two genes and the encoded amino acid sequence were analyzed by bioinformatics software. Real-time fluorescence quantitative PCR(qRT-PCR) method was used to detect the embryonic development stages of L. erythropterus(1/2 epiboly stage,blastopore closure stage,optic capsule stage,eye lens formed stage,heart-beating stage,hatching stage) and larval developmental stages(1,3,10,15 and 20 d),as well as the expression patterns in different tissues of adult fish(brain,heart,liver,muscle,black skin,red skin,stomach and retina).【Result】The full-length c DNA of RH1 gene and LWS gene of L. erythropterus were 1723 bp and 1302 bp,respectively. The RH1 gene had 465 bp 3'-UTR and 196 bp 5'-UTR with 1062 bp open reading frame(ORF),encoding 353 amino acids;the LWS gene had 213 bp 3'-UTR and 15 bp 5'-UTR with 1074 bp ORF,encoding 357 amino acids. Phylogenetic analysis showed that the two opsin genes clustered first with Perciformes,and then clustered with Killiformes,Pleuronectiformes and Cypriniformes,and finally with terrestrial vertebrates. qRT-PCR detection results showed that the relative expression level of RH1 gene was significantly higher than other periods at 15 d and 20 d after hatching(P<0.05,the same below),and the relative expression level of LWS gene was significantly higher than other periods at 20 d. The expression patterns of the two opsin genes in different tissues of adult fish were similar,and the expression levels in the retina were significantly higher than those in other tissues.【Conclusion】The expression of RH1 and LWS genes are tissue-specific,and the expression levels in early developmental stage are related to the changes of living habits of L. erythropterus,indicating that the expression of RH1 and LWS opsin genes of L. erythropterus closely corresponds to the changes of their living light environment,and plays an important role in light regulation during early developmental metamorphosis and adaptation to dark environments. |
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