黑白花奶牛白细胞介素-2基因的克隆和表达

目的]克隆牛白细胞介素-2基因(IL-2),并观察其在原核细胞中的表达。方法]应用RT-PCR技术从黑白花奶牛外周血淋巴细胞总RNA中扩增编码牛IL-2基因,并将其克隆到pGEX-2T原核表达质粒中,转化大肠杆菌BL21,经IPTG诱导后进行SDS-PAGE分析。结果]通过RT-PCR扩增获得500 bp的目的片段,所克隆的IL-2基因在原核细胞中成功表达,表达产物为分子量约43 kD的融合蛋白。结论]该研究为IL-2基因的进一步研究提供了理论依据和物质基础。

Cloning and Expression of IL-2 Gene from Holstein-Friesian Cow

Objective] The aim of this study is to clone bovine interleukin-2 gene(IL-2) and observe its expression in prokaryotic cells.Method] Bovine IL-2 gene was amplified from total RNA of peripheral blood lymphocytes of holstein-friesian cows by RT-PCR.Subsequently,the gene was cloned into pGEX-2T prokaryotic expression plasmid to construct recombinant,which were then transformed into Escherichia coli BL21.After IPTG induction,SDS-PAGE analysis was conducted.Result] A 500 bp target fragment corresponding with expectation was obtained by RT-PCR.The cloned gene successfully expressed fusion protein of about 43 kD in prokaryotic cells.Conclusion] This study provided a theoretical and material basis for further researches on IL-2 gene.