目的:克隆β2-肾上腺素能受体(2β-AR)全长基因片段。方法:根据GenBank中收录的猪β2-AR cDNA序列设计一对引物,以猪肝脏组织总RNA为模板,利用RT-PCR技术扩增目的基因,将其与pUC18载体体外连接,转化感受态大肠杆菌E.coilDH5α,筛选阳性克隆。结果:扩增出一条1257 bp的目的基因片段,该片段编码418个氨基酸。与GenBank中收录的猪2β-AR序列比对,其同源性为99.52%,编码的氨基酸有99.04%相同。结论:成功获得了β2-AR全长基因片段,为该基因的表达和受体筛药模型的建立奠定基础。
Objective: To clone the full- length gene of beta 2 - adrenergic receptor (β2 - AR). Method: Primers for PCR were designed according to the porcine ,β2 - AR eDNA sequence in the GenBank. The β2 - AR gene was amplified from the total RNA of porcine liver by RT-PCR technology, which was then cloned into pUC18 vector. The recombinant was transformed to competent cell E. coil DH5α, screening the positive clones. Result: The amplified β2 - AR gene has 1257 bases, encoding 418 amino acids. Compared with the porcineβ2 - AR DNA sequence inscribed in GenBank, the homology is 99.52% with 99.04% amino acids identieal. Conclusion: Full - length sequence of β2 - AR Gene was successfully obtained, establishing foundation for gene expression and the construction of receptor- drugs screening model.