摘要
目的 :构建携带绿色荧光蛋白 (GFP)人转化生长因子基因真核表达载体。 方法 :应用RT PCR方法扩增人转化生长因子基因 ,与GFP真核表达载体连接 ,构建带有标记基因和人转化生长因子基因的真核表达载体 ,并用限制性内切酶酶切鉴定。 结果 :构建的新质粒经酶切鉴定和DNA测序 ,证实新质粒的构建成功。 结论 :应用RT PCR方法扩增目的DNA片段 ,简单快捷 ;双酶切定向克隆可以保证构建一步到位 ,免去正反向重组体筛选的问题。
Objective: To construct an eukaryotic expression vector carring green fluorescent protein and human transforming growth factor-β_1 gene. Methods: Human transforming growth factor-β_1 gene was amplified by RT-PCR, linkaged with eGFP eukaryotic expression vector, and then identified the recombinant plasmid by restriction enzyme. Results: The recombinant plasmid carried human-TGF-β_1 and GFP gene by restriction enzyme cutting analysis and DNA sequence analysis were succeeded. Conclusion: It is simple to amplify aim gene by RT-PCR in one step. The orientation clone can assure the recombinant in one step and avoid screening obverse-inverse recombinant. The new kind of eukaryotic vector can serve as a new tool and the method in the transplantation area.
出处
《医学研究生学报》
CAS
2005年第1期13-16,共4页
Journal of Medical Postgraduates
关键词
转化生长因子
绿色荧光蛋白
真核表达
载体
Transforming growth factor-β_1
Green fluorescent protein
Eukaryotic expression
Vector
