[Efficient, stable expression of human interleukin-15 cDNA in Chinese hamster oval cells].
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.
Xue
L
L
Department of Immunology, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005.
Cui
L
L
He
W
W
Ba
D
D
D000818:Animals; D016466:CHO Cells / Q000378:metabolism; D006224:Cricetinae; D018076:DNA, Complementary / Q000096:biosynthesis* / Q000235:genetics; D019409:Interleukin-15 / Q000096:biosynthesis* / Q000235:genetics; D010957:Plasmids; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D014162:Transfection
D018076:DNA, Complementary; D019409:Interleukin-15; D012333:RNA, Messenger; D011994:Recombinant Proteins