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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
引用本文: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(3):183-192.
作者姓名:黄宝松  李金凤  张野  曹丹煜  卢晓颖  陈刚  王忠良
作者单位:广东海洋大学水产学院
基金项目:现代农业产业技术体系专项资金(CARS-47);广东海洋大学“南海学者计划”(2017年度);广东海洋大学“海之帆-起航计划”大学生科技创新培育项目(2018年度);国家级大学生创新创业训练计划项目(CXXL2018001);广东海洋大学2019年“冲一流”省财政专项资金建设项目
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2629 bp,包含1953 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

Cloning and Expression Analysis of Prolactin Receptor 1 in Rachycentron canadum at Different Water Salinities
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(3):183-192.
Authors:HUANG Bao-song  LI Jin-feng  ZHANG Ye  CAO Dan-yu  LU Xiao-ying  GHEN Gang  WANG Zhong-liang
Institution:(College of Fishery,Guangdong Ocean University,Zhanjiang 524088)
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 2629 bp,including an open reading frame of 1953 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.
Keywords:Rachycentron canadum  prolactin receptor  cDNA clone  quantitative real-time PCR  salinity adaptation
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