花鲈摄食调控相关因子全长cDNA的克隆和序列分析

Cloning and Sequence Analysis of Full-Length cDNAs of Food Intake Regulation Factors in Japanese Seabass(Lateolabrax japonicus)

本试验采用反转录聚合酶链式反应（RT-PCR）和cDNA末端快速扩增（RACE）技术克隆花鲈（Lateolabrax japonicus）调控摄食因子雷帕霉素靶蛋白（mTOR）、核糖体蛋白S6激酶beta-1（S6K1）、神经肽Y前体（NPY precursor）和脑肠肽前体（preproghrelin）cDNA全长序列。结果显示：花鲈mTOR（GenBank登录号KJ746670）cDNA全长8 296bp,开放阅读框7 551bp,编码2 516个氨基酸,预测其分子质量为286.14ku,与其他物种的相似性为62.0%～98.0%;S6 K1（GenBank登录号KJ746671）cDNA全长2 232bp,开放阅读框1 539bp,编码512个氨基酸,预测其分子质量为56.87ku,与其他物种的相似性为80.4%～84.9%;NPY precursor（GenBank登录号KJ850326）cDNA全长676bp,开放阅读框300bp,编码99个氨基酸,预测其分子质量为11.26ku,与其他物种的相似性为53.6%～99.0%;preproghrelin（GenBank登录号KJ850327）cDNA全长1 201bp,开放阅读框324bp,编码107个氨基酸,预测其分子质量为12.03ku,与其他物种的相似性为19.6%～85.0%。花鲈摄食调控因子cDNA全长序列地获得将为进一步研究其摄食调控机理奠定基础。

The full-length cDNAs of food intake regulation factors,including mechanistic target of rapamycin（mTOR）,ribosomal protein S6 kinase beta-1（S6 K1）,neuropeptide Y（NPY）precursor and preproghrelin were cloned from Japanese seabass（Lateolabrax japonicus）using reverse transcription-polymerase chain reaction（RT-PCR）and rapid amplification of cDNA end（RACE）methods.The results showed as follows：the full-length mTOR cDNA of Japanese seabass（GenBank accession No.KJ746670）was 8 296 bp,consisting of an open reading frame（ORF）of 7 551 bp encoding apolypeptide of 2 516 amino acids with a theoretical molecular weight of 286.14 ku,and shared 62.0%to 98.0%similarity with other species.The full-length S6 K1 cDNA of Japanese seabass（GenBank accession No.KJ746671）was2 232 bp,consisting of an ORF of 1 539 bp encoding apolypeptide of 512 amino acids with a theoretical molecular weight of 56.87 ku,and shared 80.4%to 84.9%similarity with other species.The full-length NPY precursor cDNA of Japanese seabass（GenBank accession No.KJ850326）was 676 bp,consisting of an ORF of 300 bp encoding apolypeptide of 99 amino acids with a theoretical molecular weight of 11.26 ku,and shared 53.6% to 99.0% similarity with other species.The full-length preproghrelin cDNA of Japanese seabass（GenBank accession No.KJ850327）was 1 201 bp,consisting of an ORF of 324 bp encoding apolypeptide of 107 amino acids with a theoretical molecular weight of 12.03 ku,and shared 19.6%to 85.0% similarity with other species.The results of this study will provide a scientific basis for further study to explain regulatory mechanism of the factors of food intake regulation in Japanese seabass.