| 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 |
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| 引用本文： | 胡骏鹏,李翔,张碧绿,范卫星,曹爱青,赵阿勇,何瑞国.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[J].中国农业科学,2008,41(1):221-228. |
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| 作者姓名： | 胡骏鹏 李翔 张碧绿 范卫星 曹爱青 赵阿勇 何瑞国 |
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| 作者单位： | 1. 华中农业人学动物科技学院,武汉,430070
2. 湖北省天门市畜牧局,湖北天门,431700 |
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| 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。
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| 关 键 词： | 朗德鹅 MSTN 肌肉组织 半定量RT－PCR IGF-I GH |
| 收稿时间： | 2006-09-08 |
| 修稿时间： | 2007-06-25 |
Cloning of MSTN Gene of Landes Goose and Research of Relationship Between MSTN Gene Expression and Different Energe Diets,or IGF-I and GH in Serum |
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| HU Jun-peng,LI Xiang,ZHANG Bi-lü,FAN Wei-xing,CAO Ai-qing,ZHAO A-yong,HE Rui-guo.Cloning of MSTN Gene of Landes Goose and Research of Relationship Between MSTN Gene Expression and Different Energe Diets,or IGF-I and GH in Serum[J].Scientia Agricultura Sinica,2008,41(1):221-228. |
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| Authors: | HU Jun-peng LI Xiang ZHANG Bi-lü FAN Wei-xing CAO Ai-qing ZHAO A-yong HE Rui-guo |
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| 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. |
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| Keywords: | Anser anser MSTN Muscle Semi-quantitative RT-PCR IGF-I GH |
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