抑制素与羊GM-CSF基因融合表达质粒pCGMISI和pEGMISI的构建

为构建抑制素基因与羊GM-CSF基因融合的真核表达质粒,通过EcoRI、XhoI酶切抑制素真核表达质粒pCISI、pEGISI及羊GM-CSF表达质粒pGM-CSF/SS,然后将GM-CSF基因分别插入pCISI、pEGISI,转化DH5α,挑选阳性克隆,抽提质粒DNA,进行酶切和测序鉴定。结果表明,构建的抑制素融合表达质粒pCGMISI和pEGMISI正确,为研究GM-CSF基因对抑制素基因疫苗的免疫增强作用奠定了基础。

Construction of inhibin expression plasmid pCGMISI and pEGMISI harboring with ovine GM-CSF

In order to construct inhibin eukaryotic expression plasmid pCGMISI and pEGISI harboring with granulocyte-macrophage colony-stimulating factor(GM-CSF),the inhibin expression plasmid pCISI,pEGISI and ovine GM-CSF plasmid pGM-CSF/SS were digested by EcoRI and XhoI,then GM-CSF gene was inserted into the large fragment including two copies of inhibin gene pCISI and pEGISI.The connected product was transferred into DH5α.A positive clone was amplified and the plasmid DNA was identified by restriction endonucleases and sequencing.The results showed that the inhibin eukaryotic expression plasmid pCGMISI and pEGMISI harboring with ovine GM-CSF had been constructed correctly,which established the foundation for enhance-immune effect of GM-CSF on the inhibin DNA vaccine.