摘要
目的构建小鼠白介素-4(murine interleukin-4,mIL-4)的真核表达载体,并研究其在293T细胞中的表达。方法 RT-PCR扩增小鼠IL-4基因后,克隆到真核表达载体pcDNA3.0上,利用电泳和测序鉴定重组质粒的大小、序列。脂质体转染法将重组质粒pcDNA3.0-mIL-4转入293T细胞中,用酶联免疫吸附法(ELISA)检测其表达。结果经酶切和测序鉴定重组质粒pcDNA3.0-mIL-4构建成功,并在293T细胞中成功表达。结论 pcDNA3.0-mIL-4可在真核细胞中成功表达,为进一步研究其在炎症性肠病(IBD)动物模型中的生物学功能提供了条件。
Objective To construct an eukaryotic expression plasmid contained routine IL-4 gene and investigate the expression of the recombinant plasmid in 293T cells. Methods The gene of routine IL-4 was obtained by RT-PCR and then was cloned into an eukaryotie expression plasmid pcDNA3.0, and then the recombinant plasmid was confirmed by electrophoresis and DNA sequencing. The plasmid was transfeeted into 293T cells by liposome transfecting technique and was identified by ELISA. Results The recombinant plasmid pcDNA3.0-raiL-4 had been constructed correctly. Meanwhile, the supernatant collecting from 293T cells, transfected with the plasmid was showed reactivity with antibody against IL-4. Conclusion The recombinant pcDNA3.0-raiL-4 was constructed successfully and highly expressed in 293T cells, which will become the basis for the further study of the biologic activity of IL-4 in vivo.
出处
《胃肠病学和肝病学杂志》
CAS
2014年第2期216-218,共3页
Chinese Journal of Gastroenterology and Hepatology
基金
广东省科技计划项目(2009B060700055)
广东省自然科学基金(10151051501000099)
