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Published on: June 10, 2015
[Efficient, stable expression of human interleukin-15 cDNA in Chinese hamster oval cells]
1Department of Immunology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005.
Objective:
To express the human interleukin 15(IL-15) cDNA in Chinese hamster oval (CHO) cells, which would benefit to the further research in the biological activities and the clinical applications of IL-15.
Methods:
The entire human interleukin-15(IL-15) coding region, deleted of all the uncoding fragments, was amplified by PCR. The amplified products were subcloned into the EcoR I and Xba I sites of the pcDNA3 plasmid, forming the recombinant eukaryotic expressing vector, pcDNA3-IL-15, which was then identified by the Bg1 II enzymatic digestion, PCR amplification and the sequence analysis. The construct was transfected into CHO cells by means of lipofectamine, followed by a series of determinations to screen the positive cell colonies, such as RT-PCR, SDS-PAGE, ELISA and CTLL-2 proliferation assay.
Results:
8 positive cell colonies highly expressed human IL-15 were obtained after G418 selection. The bioassay showed that the mean activities of the supernatants from the CHO-IL-15 cells were (318.54 +/- 32.76) U/(10(6) cells.d) and maintained stable after 6 months culture.
Conclusions:
Human IL-15 cDNA was expressed efficiently and persistently in CHO cells.
