Recombinant expression of rat and human GRO proteins in escherichia coli

Kiyoshi Konishi; Yoshimi Takata; Kazuyoshi Watanabe; Takayasu Date; Masayuki Yamamoto; Ayako Murase; Hideaki Yoshida; Tadaaki Suzuki; Susumu Tsurufuji; Motoji Fujioka

doi:10.1016/1043-4666(93)90042-4

Recombinant expression of rat and human GRO proteins in escherichia coli

Kiyoshi Konishi, Yoshimi Takata, Kazuyoshi Watanabe, Takayasu Date, Masayuki Yamamoto, Ayako Murase, Hideaki Yoshida, Tadaaki Suzuki, Susumu Tsurufuji, Motoji Fujioka

Research output: Contribution to journal › Article › peer-review

13 Citations (Scopus)

Abstract

A full-length rat gro cDNA containing the signal sequence was inserted to a plasmid/phage vector pTD-lacs which had the Escherichia coli alkaline phosphatase leader sequence downstream of the lac promoter. After removal of the gro signal sequence by site-directed mutagenesis, the vector was introduced to E. coli JM109. The cells grown in the presence of isopropyl β-D-thiogalactopyranoside were found to contain the recombinant mature rat Gro protein in the periplasmic space. The protein was released from the cells by osmotic shock, and could be purified to homogeneity from the periplasmic fluid by a single-step procedure using reverse phase high performance liquid chromatography. By similar procedures, recombinant human Groα could be obtained. In each case, about 10mg of purified cytokine were obtained from 1 litre of bacterial culture.

Original language	English
Pages (from-to)	506-511
Number of pages	6
Journal	Cytokine
Volume	5
Issue number	5
DOIs	https://doi.org/10.1016/1043-4666(93)90042-4
Publication status	Published - 1993 Sept
Externally published	Yes

Keywords

Gro/MGSA/recombinant expression/Neutrophil chemoattractant

ASJC Scopus subject areas

Immunology and Allergy
Immunology
Biochemistry
Hematology
Molecular Biology

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10.1016/1043-4666(93)90042-4

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title = "Recombinant expression of rat and human GRO proteins in escherichia coli",

abstract = "A full-length rat gro cDNA containing the signal sequence was inserted to a plasmid/phage vector pTD-lacs which had the Escherichia coli alkaline phosphatase leader sequence downstream of the lac promoter. After removal of the gro signal sequence by site-directed mutagenesis, the vector was introduced to E. coli JM109. The cells grown in the presence of isopropyl β-D-thiogalactopyranoside were found to contain the recombinant mature rat Gro protein in the periplasmic space. The protein was released from the cells by osmotic shock, and could be purified to homogeneity from the periplasmic fluid by a single-step procedure using reverse phase high performance liquid chromatography. By similar procedures, recombinant human Groα could be obtained. In each case, about 10mg of purified cytokine were obtained from 1 litre of bacterial culture.",

keywords = "Gro/MGSA/recombinant expression/Neutrophil chemoattractant",

author = "Kiyoshi Konishi and Yoshimi Takata and Kazuyoshi Watanabe and Takayasu Date and Masayuki Yamamoto and Ayako Murase and Hideaki Yoshida and Tadaaki Suzuki and Susumu Tsurufuji and Motoji Fujioka",

year = "1993",

month = sep,

doi = "10.1016/1043-4666(93)90042-4",

language = "English",

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T1 - Recombinant expression of rat and human GRO proteins in escherichia coli

AU - Konishi, Kiyoshi

AU - Takata, Yoshimi

AU - Watanabe, Kazuyoshi

AU - Date, Takayasu

AU - Yamamoto, Masayuki

AU - Murase, Ayako

AU - Yoshida, Hideaki

AU - Suzuki, Tadaaki

AU - Tsurufuji, Susumu

AU - Fujioka, Motoji

PY - 1993/9

Y1 - 1993/9

N2 - A full-length rat gro cDNA containing the signal sequence was inserted to a plasmid/phage vector pTD-lacs which had the Escherichia coli alkaline phosphatase leader sequence downstream of the lac promoter. After removal of the gro signal sequence by site-directed mutagenesis, the vector was introduced to E. coli JM109. The cells grown in the presence of isopropyl β-D-thiogalactopyranoside were found to contain the recombinant mature rat Gro protein in the periplasmic space. The protein was released from the cells by osmotic shock, and could be purified to homogeneity from the periplasmic fluid by a single-step procedure using reverse phase high performance liquid chromatography. By similar procedures, recombinant human Groα could be obtained. In each case, about 10mg of purified cytokine were obtained from 1 litre of bacterial culture.

AB - A full-length rat gro cDNA containing the signal sequence was inserted to a plasmid/phage vector pTD-lacs which had the Escherichia coli alkaline phosphatase leader sequence downstream of the lac promoter. After removal of the gro signal sequence by site-directed mutagenesis, the vector was introduced to E. coli JM109. The cells grown in the presence of isopropyl β-D-thiogalactopyranoside were found to contain the recombinant mature rat Gro protein in the periplasmic space. The protein was released from the cells by osmotic shock, and could be purified to homogeneity from the periplasmic fluid by a single-step procedure using reverse phase high performance liquid chromatography. By similar procedures, recombinant human Groα could be obtained. In each case, about 10mg of purified cytokine were obtained from 1 litre of bacterial culture.

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JO - Cytokine

JF - Cytokine

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ER -

Recombinant expression of rat and human GRO proteins in escherichia coli

Abstract

Keywords

ASJC Scopus subject areas

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