Molecular cloning of a new erythropoiesis-stimulating activity (ESA36) from Streptomyces thermoviolaceus and expression in Escherichia coli.
Previous studies have shown that an Actinomycetes strain produces a novel protein with a molecular weight of 87,000 which stimulated the growth of murine erythroid progenitors in vitro, and we purified the protein. In this study, we have isolated and sequenced a clone encoding a polypeptide which had the equivalent activity. The clone contained an open reading frame of 334 amino acids. The activity was confirmed by the expression in E. coli as a fusion protein with glutathion S-transferase (GST). Based on the differences of molecular weight and N-terminal amino acid sequences, this protein was considered to be different from that previously reported and was named "ESA36".
Sugie
M
M
Biomolecular Engineering Department, National Institute of Bioscience and Human-Technology, Ibaraki, Japan.
Oka
S
S
Tanaka
H
H
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D001483:Base Sequence; D003001:Cloning, Molecular; D003062:Codon; D004262:DNA Restriction Enzymes; D004269:DNA, Bacterial / Q000737:chemistry / Q000378:metabolism; D004920:Erythropoiesis; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics*; D008969:Molecular Sequence Data; D016366:Open Reading Frames; D010957:Plasmids; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins; D015183:Restriction Mapping; D013302:Streptomyces / Q000235:genetics*
D001426:Bacterial Proteins; D003062:Codon; D004269:DNA, Bacterial; C095987:ESA36 protein, Streptomyces thermoviolaceus; D006023:Glycoproteins; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; D004262:DNA Restriction Enzymes