γ-干扰素在NRK细胞中的表达

Expression of human γ-IFN in NRK cells

以 pBS -KS为中间载体 ,通过一步亚克隆 ,获得了两端结构为HindⅢ /XbaⅠ的γ -IFNcDNA片段 ,将该片段与真核表达载体RC/CMV进行定向重组 ,制备出γ -IFN真核表达质粒RC/CMV -IFNγ。转染NRK细胞后 ,经标准ABC法检测表明 :γ -IFN在NRK细胞中获得稳定表达。这一体系的建立为γ -IFN真核基因工程提供了稳定表达的工程细胞株 ,并为利用γ

To facilitate the insertion of γ-IFN cDNA into the eukaryotic expression vector RC/CMV, γ-IFN cDNA was subcloned from pIFNγ into the cloning vehicle pBS-KS. By this procedure, the plasmid pBS KS- IFN γ has been constructed,from which γ-IFN cDNA containing Hin dⅢ/Xba Ⅰ cohesive end can be obtained. Then the prepaired γ-IFN cNDA was ligated to the eukaryotic expression vector containing the strong CMV promotor derived from cytomegalovirus genome. γ-IFN stable eukaryotic expression system has been established. The establishment of this system can offer liable cell lines for producing recombinant γ-IFN protein, which is the same as wild γ-IFN protein;it also provides sound scientific basis for research on gene therapy and expression of γ-IFN in the mamalian cell.