一种新的番鸭MSTN基因转录本的克隆及真核表达  

Cloning and eukaryotic expression of a new MSTN gene transcription of Muscovy duck.

作　　者：郝文博[1,2] 杨晓宇[1] 李新华[1] 谷文峰[1] 胡兰[1] 

机构地区：[1]沈阳农业大学畜牧兽医学院,辽宁沈阳110161 [2]黑河学院理化系,黑龙江黑河164300

出　　处：《浙江大学学报（农业与生命科学版）》2009年第4期377-382,共6页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：辽宁省自然科学基金资助项目(20050302);辽宁省教育厅科研基金资助项目(2007304)

摘　　要：利用RT-PCR扩增MSTN cDNA全序列,然后进行克隆、测序;将重组质粒pEGFP-N1-MSTN-2通过脂质体瞬时转染番鸭成纤维细胞,用RT-PCR和SDS-PAGE检测其蛋白表达.结果表明,番鸭体内存在2种MSTN基因转录本,MSTN-1(EU336991)和MSTN-2(EU336992),大小分别为1128 bp和754 bp,同源性为94%;且MSTN-2是一个全新的MSTN转录本,含有1个开放阅读框,可编码124个氨基酸.将pEGFP-N1-MSTN-2转染番鸭成纤维细胞48 h后,在荧光显微镜下可观察到绿色荧光蛋白表达,蛋白大小约为14 kDa.这对于进一步研究MSTN基因功能及作用机理具有重要意义.The full length cDNA of MSTN was amplified by PCR, and then was cloned and sequenced. The expression protein was detected with RT-PCR and SDS-PAGE after the recombinant plasmid pEGFP-N1-MSTN-2 was transfected into fibroblasts cells using liposome method. The result shows that there were two transcriptions in Muscovy duck, MSTN-1 （EU336991） and MSTN-2 （EU336992）. MSTN-1 and MSTN-2 were 1128 bp and 754 bp separately, with homology up to 94%. MSTN-2 included an open reading frame （ORF）, encoding 124 amino acids. The green fluorescence protein was detected with fluorescence microscope at 48 h after transfection, which was about 14 kDa. This result will help to study and understand the function of MSTN gene.

关 键 词：MSTN-2 克隆 真核表达 

分 类 号：Q78[生物学—分子生物学]