牙鲆纤维蛋白原相关蛋白(FREP1)基因的克隆和表达分析  

Cloning and expression analysis of a fibrinogen-related protein(FREP1) gene from Japanese flounder(Paralichthys olivaceus)

作　　者：陈文君[1] 鲍宝龙[1] 

机构地区：[1]上海海洋大学省部共建水产种质资源发掘与利用教育部重点实验室,上海201306

出　　处：《上海海洋大学学报》2013年第4期488-495,共8页Journal of Shanghai Ocean University

基　　金：国家自然科学基金(30771668)

摘　　要：利用RACE-PCR技术获得了牙鲆FREP1(fibrinogen-relatedprotein)基因1 131 bp cDNA全长序列,其中开放阅读框786 bp,所编码的蛋白C端含有纤维蛋白样结构域标签(WWYSRCGSAGLNG)。RNA整体原位杂交检测发现只在孵化后2 d和5 d(2 dph、5 dph)仔鱼的肠道中有FREP1基因表达,而在9 dph和13 dph仔鱼胸鳍、肠道、鳃和皮肤中均有表达。荧光定量PCR检测显示该基因主要在成鱼的肠、鳃、皮肤和鳍条上表达,而脾脏、心脏、肾脏、肝脏和肌肉表达量很低或没有表达。鳍条和皮肤较其他组织均有极显著差异(P<0.01),鳃和肠相比其他组织具有显著差异(P<0.05)。由于该基因主要在鳍条、皮肤、肠和鳃中表达,提示FREP1基因可能和牙鲆先天性免疫相关。Full-length cDNA FREP1 （fibrinogen-related protein ） was cloned from larval Paralichthys olivaceus by the way of RACE-PCR, and 1 131 bp-length FREP1 cDNA with a 786 bp open reading frame encoded a putative protein containing a fibrinogen C-terminal domain signature （WWYSRCGSAGLNG）. The whole amount in situ hybridization showed that FREP1 was only expressed in intestine of 2 dph and 5 dph （days post-hatching） larvae, but expressed in fin, skin, gill and intestine of 9 dph and 13 dph larvae, qRT- PCR reveals that FREP1 was expressed mainly in fin, skin, gill and intestine of adult fish , but was expressed very lowly in liver, heart, kidney, spleen and muscle. If muscle expression was a control sample , compared with the other tissues the expression of FREP1 in fin and skin were highest （P 〈 0.01 ）. But compared with gill and intestine, the expression of FREP! in heart, kidney, spleen, liver and muscle is much lower（P 〈 0.05）. Since it was mainly expressed in skin, gill and intestine, which was associated with innate immunity system , FREP1 may be an important member of innate immune system of P. olivaceus.

关 键 词：牙鲆 纤维蛋白原相关蛋白基因 RACE-PCR 荧光定量 

分 类 号：S917[农业科学—水产科学]