中国农业科学 ›› 2008, Vol. 41 ›› Issue (1): 221-228 .doi: 10.3864/j.issn.0578-1752.2008.01.030

• 畜牧·资源昆虫 • 上一篇    下一篇

朗德鹅肌肉生成抑制因子基因(MSTN)的克隆及其表达量与日粮能量、血清IGF-I和GH的关系

胡骏鹏,李 翔,张碧绿,范卫星,曹爱青,赵阿勇,何瑞国   

  1. 华中农业大学动物科技学院
  • 收稿日期:2006-09-11 修回日期:2007-05-30 出版日期:2008-01-10 发布日期:2008-01-10
  • 通讯作者: 何瑞国

Cloning of MSTN Gene of Landes Goose and The dfference of MSTN Gene Expression by Different Energe

  

  1. 华中农业大学动物科技学院
  • Received:2006-09-11 Revised:2007-05-30 Online:2008-01-10 Published:2008-01-10

摘要: 【目的】采用反转录-聚合酶链反应(RT-PCR)技术,克隆朗德鹅肌肉生成抑制因子基因(MSTN),研究MSTN基因表达和日粮能量及其部分血清激素含量的相互关系,了解MSTN的功能,为水禽分子营养和分子育种提供基础资料。【方法】从朗德鹅(Anser anser)的腿肌中抽提总RNA,用两步法RT-PCR扩增出MSTN基因的cDNA编码序列,以pGEM-T Vector为载体,将该片段克隆到大肠杆菌(Escherichia coli)中。通过筛选阳性克隆,双酶切鉴定后测序;以MSTN基因的克隆为基础,以β-actin为内标,构建优化的半定量RT-PCR方法,研究高中低(A:13.38MJ•kg-1、B:12.13MJ•kg-1、C:10.87MJ•kg-1)3种不同能量对朗德鹅21日龄和70日龄2个时期,肌肉组织MSTN基因表达的差异;同时用放免法测定21、70日龄的血清GH和IGF-I浓度。【结果】克隆朗德鹅MSTN基因cDNA的部分序列,其片段大小为1 128bp,编码375个氨基酸组成的多肽,与已发表的鸡、鸭、鹅的MSTN 核苷酸相似性分别为94%、94%、99%;氨基酸的相似性分别为98%、97%、98%;21日龄时,朗德鹅公鹅、母鹅MSTN表达量差异不显著;70日龄时朗德鹅公鹅MSTN的表达量情况为能量A>C>B,朗德鹅母鹅MSTN的表达量情况为C>B>A;对于朗德鹅21~70日龄阶段的生长,MSTN基因的表达量和血清IGF-I的变化基本一致;与血清GH含量之间并不存在很大关联,GH和能量的高低呈正相关。【结论】日粮能量对21日龄后朗德鹅MSTN基因的表达有影响,且MSTN基因表达量和血清IGF-I具有相关性,和血清GH无较大相关性。

关键词: 朗德鹅, MSTN, 肌肉组织, 半定量RT-PCR, IGF-I, GH

Abstract: Genome RNA was extracted from muscle of Landes goose,MSTNgene mRNA was amplified by using RT-PCR.The unique DNA fragment about 1128bp in length was obtained and the PCR product was cloned into pGEM-T vector. The MSTN gene was isolated and sequenced from the positive clones screened.Sequence analysis revealed that it has a length of 1128 nucleotides which encodes a 375 amino peptide. Compared with homologous sequence of human, rat,duck and goose,it displayed a fairly high degree conservation.The homologue of the nucleotide sequence are 94%,94%,99% respectively;the homologue of the amino acid sequence are 98%,97%,98% respectively. Based on the MSTN gene clone, using β-actin as inner control, constructing an optimal semi-quantitative RT-PCR method,the difference of MSTN gene expression at 21 days and 70days of Landes goose by feeding different energe diets was researched ,in the same time we detected IGF-I and GH in serum. The results showed the expressions of MSTN in female landes geese were a little higher than male at 21 days;but the expression of MSTN in male was A>C>B by 70d,to female was C>B>A by 70d. In 21~70 days the expression of MSTN was changed with IGF-I in serum;it was not associated with GH in serum,GH was significant correlations with energy.

Key words: Anser anser, MSTN, Muscle, Semi-quantitative RT-PCR, IGF-I, GH