Synthesis and secretion of human nerve growth factor by Saccharomyces cerevisiae.
The DNA coding for human nerve growth factor (hNGF) was chemically synthesized and introduced into Saccharomyces cerevisiae. Expression and secretion of hNGF was obtained by use of the yeast phosphoglycerate kinase-encoding gene promoter and the pre-pro sequence of the yeast alpha-mating factor. Immunoblotting with antiserum raised against a protein A-hNGF fusion protein, allowed the detection of an immunoreactive material secreted into the culture medium. A preparation from the culture medium, partially purified by ion-exchange column chromatography, stimulated neurite outgrowth from rat pheochromocytoma PC12h cells.
Kanaya
E
E
Mitsubishi Kasei Institute of Life Sciences, Tokyo, Japan.
Higashizaki
T
T
Ozawa
F
F
Hirai
K
K
Nishizawa
M
M
Tokunaga
M
M
Tsukui
H
H
Hatanaka
H
H
Hishinuma
F
F
D000595:Amino Acid Sequence; D001483:Base Sequence; D015153:Blotting, Western; D015870:Gene Expression / Q000502:physiology; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009414:Nerve Growth Factors / Q000096:biosynthesis / Q000235:genetics*; D009710:Nucleotide Mapping; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D011995:Recombination, Genetic; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism
D009414:Nerve Growth Factors; D011994:Recombinant Proteins