山羊SPRN基因真核表达载体的构建

Construction of Eukaryotic Expression Vector of Caprine SPRN Gene

以山羊小脑cDNA为模板,利用RT-PCR法扩增获得了SPRN基因完整编码区片段,将其连接到pMD19-T载体上,转化感受态JM109后,筛选并鉴定阳性克隆,通过序列测量,将序列正确的片段双酶切后连接到真核表达载体pEGFP-N1上。结果表明,成功构建了山羊SPRN基因完整编码区的真核表达载体。

Using the cDNA of caprine cerebellum as templates, the full-length open reading frame of SPRN gene was obtained by using RT-PCR and ligated to vector pMD19-T for JM109 transformation. After screening and identifying, the positive clone was sequenced and ligated to eukaryotic expression vector pEGFP-N1 after double digestion. The results confirmed that the eukaryotic expression vector of caprine SPRN coding regions was constructed successfully.