【作者】 王晓玉； 徐黎明； 吴秀梅； 纪锋； 赵景壮； 刘淼； 曹永生； 卢彤岩； 尹家胜；

【Author】 WANG Xiaoyu;XU Liming;WU Xiumei;JI Feng;ZHAO Jingzhuang;LIU Miao;CAO Yongsheng;LU Tongyan;YIN Jiasheng;Heilongjiang River Fishery Research Institute Chinese Academy of Fishery Sciences;

【机构】 中国水产科学研究院黑龙江水产研究所； 上海海洋大学水产与生命学院；

【摘要】 胰岛素样生长因子-I(Insulin-like growth factor-I,IGF-I)在鱼类的生长发育过程中起着重要的调节作用。此研究采用逆转录聚合酶链式反应(RT-PCR)方法,从哲罗鲑肝脏的总RNA中扩增出IGF-I的c DNA开放阅读框(Open reading frame,ORF)序列,运用软件对其进行生物信息学分析,并利用荧光实时定量PCR技术检测了哲罗鲑成鱼不同组织中IGF-I m RNA的表达情况。结果显示,IGF-I基因的c DNA开放阅读框为573 bp,编码190个氨基酸,蛋白质等电点为9.21,氨基酸结构由信号肽、B、C、A、D结构域及E肽组成;氨基酸序列与其他鲑科鱼类具有较高的同源性,其中与北极红点鲑的IGF-I同源性最高(99.2%);组织表达分析显示,哲罗鲑IGF-I m RNA在肝脏中表达量最高,在鳃、前肠中次之,在脑、头肾、脾、心、胃和肌肉等组织中的表达量较低。此研究为鱼类IGF-I基因功能研究奠定基础,为哲罗鲑生长调控提供科学依据。更多还原

【Abstract】 Insulin-like growth factor-I(IGF-I) plays a major role in the control of growth and development in fish. Total RNA was isolated from Huchotaimen liver tissue. The c DNA encoding IGF-I peptide was amplified by reverse transcription polymerase chain reaction(RT-PCR) strategy using isolated total RNA as template. The IGF-I m RNA level was detected in different tissues using real-time quantitative PCR technique. The results showed that the IGF-I gene c DNA contains a open reading frame(ORF) of 573 bp, encoding a polypeptidewith length of 190 amino acids and isoelectric point of 9.21. The polypeptide is composed of siginal peptide, B, C, A, D domain and E peptide. The amino acid sequence of Hucho taimen IGF-I was high homologus to that of other Salmonidae. Hucho taimen shared the highest identity with Salvelinus alpinus and the homology at nucleotide level of the IGF-I gene were 99.2%. The expression analysis of IGF-I gene with real-time quantitative RT-PCR showed that the highest level of IGF-I m RNA was observed in liver, followed in gill and anterior intestine. There was no obvious expression in brain, pronephros, spleen, heart, stomach and muscle.更多还原