Biologically active human and mouse nerve growth factors secreted by the yeast Saccharomyces cerevisiae.
Nerve growth factor (NGF) is a trophic agent that is essential for the development and survival of sympathetic and sensory nerves. A chemically-synthesized DNA fragment encoding human NGF (hNGF) and a cDNA encoding mouse NGF (mNGF) were engineered for expression in the yeast, Saccharomyces cerevisiae. Expression and secretion of hNGF and mNGF was attempted under the direction of the yeast PGK promoter and with various leader sequences. Among the leader sequences tested, that of the yeast alpha-factor successfully directed secretion of both hNGF and mNGF that were correctly processed. The content of the recombinant NGF (reNGF) in the culture supernatant was estimated to be 1 microgram/ml. The yeast-produced reNGF was able to bind to NGF receptors in rat pheochromocytoma (PC12) cells as efficiently as the standard mNGF, and partially purified reNGF could induce neurite outgrowth of PC12 cells. Thus, we have demonstrated that biologically active human and mouse reNGF can be produced in yeast cells.
Nishizawa
M
M
Laboratory of Molecular Genetics, Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Ozawa
F
F
Higashizaki
T
T
Hirai
K
K
Hishinuma
F
F
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001709:Biotechnology; D002460:Cell Line; D004274:DNA, Recombinant / Q000235:genetics; D005818:Genetic Engineering; D005822:Genetic Vectors; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D016501:Neurites / Q000187:drug effects / Q000648:ultrastructure; D012441:Saccharomyces cerevisiae / Q000235:genetics
D004274:DNA, Recombinant; D009414:Nerve Growth Factors