Characterization of a beta-nerve growth factor expression vector for mammalian cells.
A mammalian expression vector that directs expression of murine beta-nerve growth factor (beta-NGF) from a murine sarcoma virus long terminal repeat (LTR) promoter element was constructed and characterized. The vector, designated pLTRSNGF, was stably transfected into murine L-cells, and beta-NGF mRNA and protein levels were quantified and compared to endogenous levels in control L-cells. Transfection of pLTRSNGF resulted in an approximate doubling of both beta-NGF mRNA and mature beta-NGF protein secreted into the media. Transfection of pLTRSNGF into rat PC 12 cells resulted in colonies of autocrine-differentiating cells that extended dense networks of neurites in the absence of added NGF, indicating that the beta-NGF produced from the vector is biologically active.
Clegg
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Neuroscience Research Institute, University of California, Santa Barbara 93106.
D000818:Animals; D002478:Cells, Cultured; D005822:Genetic Vectors; D007739:L Cells; D051379:Mice; D009414:Nerve Growth Factors / Q000235:genetics* / Q000378:metabolism; D016716:PC12 Cells; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012333:RNA, Messenger / Q000378:metabolism; D012091:Repetitive Sequences, Nucleic Acid; D009053:Sarcoma Viruses, Murine / Q000235:genetics; D014162:Transfection
D009414:Nerve Growth Factors; D012333:RNA, Messenger