牙鲆lin-29基因的全长cDNA克隆、鉴定及其在早期发育中的表达分析

通过PCR技术,从牙鲆(Paralichtys olvaceus)总RNA中获得牙鲆lin-29基因c DNA序列,该序列全长1 769 bp,包括1 329 bp开放阅读框,编码442个氨基酸,预测其是一种多锌指结构蛋白;氨基酸序列对比分析显示,牙鲆LIN-29与大黄鱼(Larimichthys crocea)的同源性最高,达95.25%;系统进化树分析表明,牙鲆与其他鱼类聚集一支,并且与罗非鱼的进化关系最近。荧光定量PCR显示,在牙鲆胚胎和仔鱼发育阶段,以原肠胚期lin-29表达量最高;组织表达分析表明,脑中表达最高,肌肉中最低。外源甲状腺激素(TH)在牙鲆仔鱼发育初期(TH处理后2 d、5 d)下调lin-29表达,TH处理后21 d(36 dph)和26 d(41 dph)上调lin-29基因表达,同时,硫脲(TU)处理均显著上调lin-29表达。结果显示牙鲆lin-29基因可能在牙鲆早期发育过程中扮演着重要的角色,为进一步研究lin-29在牙鲆生长发育中的功能奠定基础。

Full length cDNA cloning, identification and expression analysis of lin-29 during early development of Japanese flounder, Paralichtys olivaceus

Lin-29 gene from Japanese flounder (Paralichthys olivaceus) was cloned by PCR, and its gene sequence is 1 769 bp including a 1 329 bp open reading frame (ORF) which encodes a polypeptide of 442 amino acids. The lin-29 gene was predicted encoding a zinc finger protein and was highly similar to amino acid sequence in other fish through phylogenetic analysis. BLAST analysis indicated that the lin-29 from P. olivaceus showed an average homology of 95.25 % to Larimichthys crocea. Real-time quantitative PCR revealed that expression level of the lin-29 was the highest during gastrulation stage and in the tissue of brain. The expression of lin-29 was highly up or down-regulated by thyroid hormone (TH) at different stages while thiourea (TU) up-regulated significantly the lin-29 levels. The results showed that LIN-29 played an important role in development and metabolism of P. Olivaceus. These results provided the basic information for further study on the role of lin-29 gene in the development of P. olivaceus.