军曹鱼催乳素受体PRLR1基因的克隆及其在不同盐度条件下mRNA 表达差异

doi:10.13560/j.cnki.biotech.bull.1985.2019-0813

摘要/Abstract

摘要： 催乳素受体通过结合催乳素,能调节鱼体渗透压。为研究催乳素受体1（PRLR1）在高盐水体和低盐水体中对军曹鱼（Rachycentron canadum）的渗透调节作用,利用cDNA末端快速扩增（RACE-PCR）技术,获得了军曹鱼PRLR1全长cDNA序列。该基因全长为2 629 bp,包含1 953 bp的开放阅读框ORF,可编码650个氨基酸。氨基酸序列包含了2个纤维连接蛋白3型结构域（FN3）、保守的WS区和box1。采用qRT-PCR技术,检测不同盐度（10‰、30‰和35‰）条件下鳃、肠、体肾中PRLR1基因mRNA表达情况。结果显示,PRLR1基因在军曹鱼的各个组织中均有表达,其中鳃表达量最高,其次是肌肉、体肾和肠,而在胃、脾、脑和心脏中则微量表达。低盐组、正常组和高盐组中,PRLR1基因的表达量均为鳃最高;肠次之;体肾最低。随着盐度提高,PRLR1基因的鳃、肠和体肾组织表达量变化规律均呈逐步下降趋势。以上结果反映了军曹鱼PRLR1在渗透压器官中的功能差异性,说明PRLR1在军曹鱼渗透压调节上具有重要作用。

关键词: 军曹鱼, 催乳素受体, cDNA克隆, 荧光定量PCR, 盐度适应

Abstract: Prolactin receptor can regulate osmotic pressure of fish by binding prolactin. In order to understand the expression of prolactin receptor PRLR1 in cobia（Rachycentron canadum）in high saline and low saline,the full-length cDNA sequence of codfish PRLR1 was obtained by RACE-PCR（rapid amplification of cDNA ends）. The full-length cDNA of this gene was 2 629 bp,including an open reading frame of 1 953 bp,and encoding 650 amino acids. The amino acid sequence comprised of two fibronectin type 3 domains（FN3）,a conserved WS region,and box1. The expression of PRLR1 mRNA in gill,intestine and body kidney was detected by real-time quantitative PCR under different water salinities（10‰,30‰,and 35‰）. The results showed that gene PRLR1 was expressed in various tissues of cobia,among which the highest expression of gill,followed by muscle,body kidney and intestine,but slightly expressed in stomach,spleen,brain and heart. In the low-salt group,the normal group and the high-salt group,the expression level of gene PRLR1 in gill was the highest,followed by that in the intestine and the lowest in the kidney. With the increase of salinity,the expression of PRLR1 in gill,intestine and body kidney showed a gradual decline. The above results reflect the functional difference of PRLR1 in osmotic organs of R. canadum,indicating that PRLR1 plays an important role in osmotic pressure regulation.

Key words: Rachycentron canadum, prolactin receptor, cDNA clone, quantitative real-time PCR, salinity adaptation

黄宝松, 李金凤, 张野, 曹丹煜, 卢晓颖, 陈刚, 王忠良. 军曹鱼催乳素受体PRLR1基因的克隆及其在不同盐度条件下mRNA 表达差异[J]. 生物技术通报, 2020, 36(3): 183-192.

HUANG Bao-song, LI Jin-feng, ZHANG Ye, CAO Dan-yu, LU Xiao-ying, GHEN Gang, WANG Zhong-liang. Cloning and Expression Analysis of Prolactin Receptor 1 in Rachycentron canadum at Different Water Salinities[J]. Biotechnology Bulletin, 2020, 36(3): 183-192.

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