【摘要】 目的克隆Exendin-4 39肽氨基酸序列基因编码区cDNA。方法根据Exendin-4氨基酸序列编码cDNA,设计正、负向引物/模板链,利用重叠延伸PCR法扩增出Exendin-4编码区基因DNA片段;将获得的基因片段插入pGEM-T-Easy质粒中;转化到大肠杆菌DH5α后挑选阳性克隆,利用限制性内切酶及核苷酸序列分析技术鉴定重组质粒。结果经质粒DNA酶切分析及序列测定,获得了Exendin-4 DNA片段序列。结论本研究成功克隆了Exendin-4 cDNA,为下一步构建Exendin-4真核表达载体打下基础,也为Exendin-4基因治疗糖尿病提供了前提条件。更多还原

【Abstract】 Objective To clone and identity the Exendin-4 cDNA of 39 peptides amino acid sequences of the genes encoding area.Methods The forward and reverse primer were designed according to the Exendin-4 cDNA by comple mentary primer/template PCR.The Exendin-4 open reading frame(ORF) was cloned by overlap extension PCR.The Exendin-4 cDNA fragment was inserted in pGEM-T-Easy vector after digested with restriction enzyme and DNA sequencing.The positive cloning was chosen after the recombinant vectors transforming to Escherichia coli DH5 α.Recombinant plasmid was identified by restriction enzymes and nucleotide sequence analysis technology.Results The Exendin-4 cDNA clone was confirmed by restriction enzyme digestion and DNA sequencing.Conclusion The Exendin-4 cDNA clone is constructed successfully in this study.It provide biases for constructing Exendin-4 eukaryotic expression vector and diabetes gene therapy.更多还原