牙鲆纤维蛋白原相关蛋白(FREP1)基因的克隆和表达分析
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国家自然科学基金(30771668)


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

    利用RACE-PCR技术获得了牙鲆FREP1(fibrinogen related protein)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。

    Abstract:

    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 FREP1 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.

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陈文君,鲍宝龙.牙鲆纤维蛋白原相关蛋白(FREP1)基因的克隆和表达分析[J].上海海洋大学学报,2013,22(4):488-495.
CHEN Wen-jun, BAO Bao-long. Cloning and expression analysis of a fibrinogen related protein(FREP1)gene from Japanese flounder(Paralichthys olivaceus)[J]. Journal of Shanghai Ocean University,2013,22(4):488-495.

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  • 在线发布日期: 2013-08-15
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