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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。
The full-length 1 131 bp cDNA of FREP1 gene was obtained by RACE-PCR. The open reading frame (ORF) was 786 bp in length. The encoded protein contained a fibrin-like domain tag (WWYSRCGSAGLNG) at its C-terminus. RNA whole in situ hybridization assay showed that FREP1 gene expression was found only in larvae of larvae at 2 d and 5 d after hatching (2 dph, 5 dph), while in larval fin, intestine, gill and skin of 9 dph and 13 dph larvae Express. Fluorescent quantitative PCR showed that the gene was mainly expressed in intestine, gill, skin and ray of adult fish, while the expression of spleen, heart, kidney, liver and muscle was very low or not expressed. There was significant difference between fin and skin in other tissues (P <0.01), but there was significant difference between gill and intestine in other tissues (P <0.05). As the gene is mainly expressed in the rays, skin, intestine and gills, suggesting that FREP1 gene may be associated with innate immunity of Japanese flounder.