CAJ | 학술논문

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.
이용RT-PCR확증 MSTN cDNA전서렬,연후진행극륭、측서;장중조질립 pEGFP-N1-MSTN-2통과지질체순시전염번압성섬유세포,용RT-PCR화SDS-PAGE검측기단백표체.결과표명,번압체내존재2충MSTN기인전록본,MSTN-1(EU336991)화MSTN-2(EU336992),대소분별위1128 bp화754 bp,동원성위94%;차MSTN-2시일개전신적MSTN전록본,함유1개개방열독광,가편마124개안기산.장pEGFP-N1-MSTN-2전염번압성섬유세포48 h후,재형광현미경하가관찰도록색형광단백표체,단백대소약위14 kDa.저대우진일보연구MSTN기인공능급작용궤리구유중요의의.
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.