摘要
目的构建重组小鼠干扰素γ(mIFN-γ)真核表达系统,并检测其在cos-7细胞中的表达。方法用RTPCR方法,从ConA活化的小鼠脾细胞中扩增出mIFNγcDNA,将其克隆至pGEMT载体上,测序正确后,再定向连接到真核表达载体pcDNA3中,经DEAEDextran转染至cos7细胞,检测其在细胞内外的表达。结果RTPCR扩增的mIFNγcDNA与GenBank中mIFNγ序列一致。构建的真核重组表达质粒pcDNA3/mIFNγ转染cos7细胞后,可检测到mIFN-γ的转录和表达。结论已成功构建了mIFNγ真核表达系统,为抗病毒基因疫苗的研究奠定了基础。
Objective To construct an eukaryotic expression system and express recombinant murine IFN-γ（mIFN-γ）in cos-7 cells. Methods Amplify mIFN-γ cDNA from ConA-activated murine splenocytes by RT-PCR and clone into pGEM-T vector. Identify the recombinant plasmid by sequencing and directly ligate to eukaryotic expression vector pcDNA3, then transfect cos-7 cells in the mediation of DEAE-Dextran and determine the intracellular and extracellular expressions of mIFN-γ. Results The sequence of amplified mIFN-γ cDNA was identical to that reported in GenBank. The transcription and expression of mlFN-γ was confirmed in the cos-7 cells transfected with pcDNA3/mIFN-γ. Conclusion The eukaryotic expression system of mIFN-γ was successfully constructed. It laid a foundation of development of antivirus gene vaccine.
出处
《中国生物制品学杂志》
CAS
CSCD
2006年第3期265-267,共3页
Chinese Journal of Biologicals
基金
河北省自然科学基金资助项目(2004000631).
