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  

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

作　　者：高丽娜[1] 付元帅[1] 施志仪[1] 苏艳芳[1] 张红梅[1] 

机构地区：[1]上海海洋大学农业部淡水水产种质资源重点实验室,上海201306

出　　处：《上海海洋大学学报》2016年第1期8-17,共10页Journal of Shanghai Ocean University

基　　金：国家自然科学基金(31172392);上海市教育委员会科研创新项目(1522083);上海市自然科学基金(15ZR1420600)

摘　　要：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。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 fishes 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.

关 键 词：lin-29 牙鲆 变态发育 甲状腺激素 硫脲 基因表达 

分 类 号：S917[农业科学—水产科学]