家禽MSTN基因转录子的克隆及真核表达  

Cloning and eukaryotic expression of MSTN gene transcripton in chick ens

作　　者：胡兰[1] 隋世慧[1] 

机构地区：[1]沈阳农业大学畜牧兽医学院,沈阳110161

出　　处：《东北农业大学学报》2012年第12期97-103,共7页Journal of Northeast Agricultural University

基　　金：辽宁省自然科学基金资助项目(20090302);辽宁省教育厅科研基金资助项目(2007304)

摘　　要：为了确定家禽中MSTN基因转录子的种类、序列及表达差异,对1日龄海兰鸡、白羽鸡和寿光鸡腿肌MSTN基因进行RT-PCR扩增、TA克隆、测序及SDS-PAGE分析。结果表明,海兰鸡、白羽鸡和寿光鸡鸡腿肌中均存在3种不同大小的MSTN cDNA;cMSTN-1由1 128 bp组成,海兰鸡与白羽鸡的cMSTN的序列相同,寿光鸡在第1 115碱基处由G变为A;cMSTN-2由985 bp组成,缺失了cMSTN-1的374～517 bp处序列;海兰鸡和白羽鸡的cMSTN-2与寿光鸡的cMSTN-2相比,均有5处碱基不同,而海兰鸡和白羽鸡只存在2处碱基不同;cMSTN-2在388～390 bp处出现UAA终止密码子;cMSTN-3由754 bp组成,缺失了374～748处的374个核苷酸,在上述三种家禽中cMSTN-3序列相同。转染成纤维细胞48 h后,pEGFP-N1-cMSTN-2表达蛋白质分子质量约为15 ku,而pEGFP-N1-cMSTN-3不表达。这是首次发现三个品种的鸡体内同时存在三种不同大小的MSTN基因转录子,该结果可为进一步确定MSTN基因的作用机理提供试验依据。In order to investigate the kinds, sequences and expression difference of MSTN transcription in chickens, RT-PCR, TA cloning and sequencing was applied in this experience. The results showed that there were three MSTN gene transcriptons of varied sizes in leg muscle of Hailan, Baiyu and Shouguang chicken aged 1 day. cMSTN-1 was composed of 1 128 bp, and Hailan chicken and Baiyu chicken had the same sequences, but Shouguang chicken had G→A at 1 115 bp. cMSTN-2 was consisted of 985 bp, which deleted 143 cases from 374 to 517 of cMSTN-1, cMSTN-2 of Hailan and Baiyu chickens had 5 bp difference from Shouguang chicken, but only 2 bp difference each other. There was UAA in 388-390 bp of cMSTN-2, cMSTN-3 was composed of 754 bp, and 374 cases from 374 to 748 were deleted compared to that of cMSTN-1, which was the same in the above three kinds of chickens. pEGFP-NI-cMSTN-2 and pEGFP-NI-cMSTN-3 were transfected into fibroblasts using liposome method. for 48 h, pEGFP-NI-cMSTN-2 produced protein which was about 15 ku. In this report, we presented the first biochemical evidence to show that there were different MSTN cDNA in chickens, which provided experimental basis to study the mechanism of MSTN gene in further research.

关 键 词：MSTN基因 转录子 家禽 克隆 真核表达 

分 类 号：S858.31[农业科学—临床兽医学] Q78[农业科学—兽医学]