PcDNA3.1-rhGM-CSF重组质粒的构建及鉴定

Construction and Identification of Eukaryotic Expression Vector of PcDNA3.1-rhGM-CSF

目的:构建PcDNA3.1-rhGM-CSF真核细胞表达载体。方法:采用PCR、T-A克隆及基因定向克隆技术,将rhGM-CSF基因插入PcDNA3.1(+)空载体,构建了PcDNA3.1-rhGM-CSF真核表达载体。经限制性内切酶酶切和DNA测序进行鉴定。结果:双酶切及DNA测序证实PcDNA3.1-rhGM-CSF真核细胞表达载体构建成功。结论:成功构建PcDNA3.1-rhGM-CSF真核细胞表达载体。

Objective：To construct PcDNA3.1-rhGM-CSF expression vector in order to make clear the role of rhGM-CSF in the differentiation of leukemic cell.Methods： The eukaryotic expression plasmid PcDNA3.1-hGM-CSF was constructed by means of PCR,T-A clone and directional cloning techniques.The combinants were finally sequenced and identified by restriction endonuclease digestion.Results： PcDNA3.1-rhGM-CSF expression vector was successfully constructed and identified by double endonuclease digestion and sequence analysis.Condusion： PcDNA3.1-rhGM-CSF expression vector has been successfully constructed.