Renaturation, purification, and characterization of human truncated macrophage colony-stimulating factor expressed in Escherichia coli.

Renaturation, purification, and characterization of human truncated macrophage colony-stimulating factor expressed in Escherichia coli. Research Abstract Details 

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Renaturation, purification, and characterization of human truncated macrophage colony-stimulating factor expressed in Escherichia coli. Abstract Text:

K Yamanishi,M Takahashi,T Nishida,Y Ohmoto,M Takano,S Nakai,Y Hirai,

A human truncated macrophage colony-stimulating factor (M-CSF) encoding the amino acid residues from 3 to 153 of the native M-CSF was expressed by using a two-cistron expression system in Escherichia coli. The truncated M-CSF found in inclusion bodies was renatured and had CSF activity. Purification, which included a QAE-ZeTa preparative cartridge concentration step followed sequentially by HPLC on TSK-gel Phenyl-5PW and TSK-gel DEAE-5PW columns, gave an overall yield of 63.8%. The purified truncated M-CSF had a specific activity of 4 x 10(7) units/mg of protein. Peptide mapping of a lysylendopeptidase digest by reversed-phase HPLC confirmed the amino acid sequence predicted from the cDNA sequence. SDS-PAGE of the purified truncated M-CSF gave a single band at 17 kDa under reducing conditions and at 32 kDa under non-reducing conditions. Activated Thiol-Sepharose 6B column chromatography and other experiments failed to detect any free cysteine residue in spite of the existence of 7 cysteine residues in the truncated M-CSF subunit. These results indicate that it is a dimeric structure linked by one or more intermolecular disulfide bonds.

Renaturation, purification, and characterization of human truncated macrophage colony-stimulating factor expressed in Escherichia coli. Publishing Authors By Initials

K Yamanishi,M Takahashi,T Nishida,Y Ohmoto,M Takano,S Nakai,Y Hirai,

For similar investigative techniques: genetic techniques: gene transfer techniques: transfection research abstracts see: investigative techniques: genetic techniques: gene transfer techniques: transfection research

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Renaturation, purification, and characterization of human truncated macrophage colony-stimulating factor expressed in Escherichia coli. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Journal of biochemistry

VOLUME: 109

Page Numbers: 404-9

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Mar

YEAR: 1991

Renaturation, purification, and characterization of human truncated macrophage colony-stimulating factor expressed in Escherichia coli. Information

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LANGUAGE: eng

NlmUniqueID: 376600

Renaturation, purification, and characterization of human truncated macrophage colony-stimulating factor expressed in Escherichia coli. Keywords Mesh Terms:

KEYWORDS: Transfection

MESH TERMS: chemistry

Chemical & Substance for Abstract: Renaturation, purification, and characterization of human truncated macrophage colony-stimulating factor expressed in Escherichia coli. Information

Substance Name: Macrophage Colony-Stimulating Factor

Registry Number: 81627-83-0

Grant and Affiliation Information for Renaturation, purification, and characterization of human truncated macrophage colony-stimulating factor expressed in Escherichia coli.

AFFILIATION: Cellular Technology Institute, Otsuka Pharmaceutical Co., Ltd., Tokushima.

Country: JAPAN

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MEDLINETA: J Biochem

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