CAJ | 학술논문

[目的]克隆牛的卵泡抑素基因（Follistatin,FSTN）基因,构建真核表达载体。[方法]用Trizol法从牛的卵巢中提取总RNA,反转录成cDNA,用带有酶切位点牛FSTN的特异性引物扩增其完整编码区序列,连接到T载体、测序,序列无误后亚克隆入真核表达载体pIRES2-AcGFP1中,酶切及PCR鉴定载体。[结果]经酶切及PCR鉴定表明成功构建了真核表达载体pIRES2-AcGFP1-FSTN。[结论]成功构建了真核表达载体pIRES2-AcGFP1-FSTN,为促进肌肉生长的转基因优质肉牛品种培育奠定了基础。
[목적]극륭우적란포억소기인（Follistatin,FSTN）기인,구건진핵표체재체。[방법]용Trizol법종우적란소중제취총RNA,반전록성cDNA,용대유매절위점우FSTN적특이성인물확증기완정편마구서렬,련접도T재체、측서,서렬무오후아극륭입진핵표체재체pIRES2-AcGFP1중,매절급PCR감정재체。[결과]경매절급PCR감정표명성공구건료진핵표체재체pIRES2-AcGFP1-FSTN。[결론]성공구건료진핵표체재체pIRES2-AcGFP1-FSTN,위촉진기육생장적전기인우질육우품충배육전정료기출。
【Objective】To clone and construct the eukaryotic expression vector containing the bovine follistatin cDNA sequence.【Methods】Total RNA was extracted from bovine ovary by Trizol total RNA Extract Kit,and the whole coding region of bovine follistatin cDNA gene was cloned by RT-PCR using specific primers containing restriction enzyme cutting site.The purified bovine follistatin cDNA was inserted into pMD18-T vector and was then sequenced,then subclone the correct follistatin cDNA to eukaryotic expression vector pIRES-AcGFP.Double-enzyme cleavage and PCR test were used to identify the vector.【Results】Double-enzyme cleavage and PCR test showed that FSTN was successfully cloned into eukaryotic expression vector.【Conclusion】We successfully constructed the eukaryotic expression vector pIRES2-AcGFP1-FSTN.This study provided the foundation for fostering high beef quality transgenic cattle of promoting muscle growth.