真核表达质粒pcDNA3.1a-FGFR1构建及意义

Construction of eukaryotic expression plasmid pcDNA3.1a-FGFR1 and its significance

目的:构建真核表达质粒pcDNA3.1a-FGFR1。方法:通过PCR扩增FGFR1目的片段,双酶切纯化PCR产物及pcDNA3.1a,将扩增的FGFR1基因片段插入pcDNA3.1a线性质粒,即构建成pcDNA3.1a-FGFR1真核表达质粒,将质粒转化感受态DH5α菌株,筛选阳性克隆行双酶切鉴定及测序鉴定。结果:酶切及测序结果表明pcD-NA3.1a-FGFR1真核表达质粒构建成功。结论:pcDNA3.1a-FGFR1真核表达质粒构建成功,为下一步对心血管疾病研究奠定了一定的实验基础。

Objective： To construct a recombinant eukaryotic expression plasmid pcDNA3.1a-FGFR1.Methods： The fragment of FGFR1 was amplificated by PCR,after purification,the PCR products of FGFR1 and eukaryotic expression plasmid pcDNA3.1a were cut and FGFR1 were inserted into pcDNA3.1a,and then transformed into DH5α strain and positive clone was selected.The recombinant plasmid was confirmed by restriction endonuclease analysed and sequencing test.Results： DNA sequencing and restriction endonuclease digestion analysis indicated that the eukaryotic expression plasmid pcDNA3.1a-FGFR1 was constructed successfully.Conclusion： Construction of pcDNA3.1a-FGFR1 eukaryotic expression plasmid was successful.This study for the next step on angiocardiopathy research laid the experimental foundation.