ABSTRACT
Objective:To identify the expression of egfp-hri fusion on NIH3T3 cells which is carried by recombinant retrovirus. Methods:The egfp-hri fusion gene was integrated intoNIH3T3 cells by the infection of recombinant viral supernatant from the G418 resistent PA317 cells transfected stably with the plasmid of pLNCX-EGFP-C1-hri. The expression effect of egfp-hri fusion was visualized under the fluorescent microscope, while the expression levels of egfp-hri fusion protein by Western blot. Results:The observation from fluorescent microscope showed the green fluorescent was obviously observed in the cytoplasm , western-blotting showed the expression level of egfp-hri fusion gene increased by 83%and 81%, as compared with those uninfected cells and in the cells infected with empty vector, respectively. Conclusion:The infected NIH3T3 cells can express egfp-hri fusion protein successfully.
ABSTRACT
Objective:To identify the expression of plasmid pLNCX-EGFP-C1-hri targeting the gene of Human ribonuclease inhibitor (hri) in PA317 cells which is capable of expression in mammalian cells.Methods: The vector of pLNCX-EGFP-C1-hri was transfected into PA317 cells by Lipofectamine 2000 and then the expression of recombinated plasmid was verified in living cells by observing the transcription level of egfp-hri fusion gene mRNA with RT-PCR method and the expression level of egfp-fusion hRI protein with western blotting method respectively.Results: Both RT-PCR and western blotting showed the egfp-hri fusion gene was obviously expression in PA317 cells.Conclusion: The plasmid of pLNCX-EGFP-C1-hri targeting hRI is successfully constructed and the protein of hRI can be expressed in PA317 cells correctly.