【摘要】 目的构建重组鼠白细胞介素2的真核表达体系,获得mIL2基因的表达质粒,用其提高DNA疫苗的免疫原性。方法从经PMA刺激的小鼠脾细胞中提取总RNA,采用RTPCR技术调取鼠白细胞介素2基因,并将其克隆至真核表达质粒VR1012中,转染COS7细胞,将转染细胞裂解,进行Westernblot检测。结果获得完整的带有信号肽的mIL2序列的质粒,经Westernblot检测阳性。结论构建了mIL2真核表达体系,并获得有生物活性的mIL2蛋白。更多还原

【Abstract】 Objective To construct an eukaryotic expression system for recombinant murine interleukin-2(mIL-2) and use the recombinant plasmid as an adjuvant to enhance the immunogenicity of DNA vaccine.Methods Extract total RNA from murine splenocytes immunized with PMA and amplify mIL-2 gene by RT-PCR.Clone the amplified gene into an eukaryotic expression vector VR1012 and transfect to COS-7 cells.Split the transfected cells and identify the expressed product by Western blot.Results A mIL-2 gene sequence with signal peptide was obtained and inserted into plasmid VR1012 correctly.Western blot proved that mIL-2 was expressed.Conclusion An eukaryotic expression system for mIL-2 was constructed,and mIL-2 with biological activity was obtained.It laid a foundation of large-scale production of mIL-2 protein.更多还原