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 journalArticlepeer-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 languageEnglish
Pages (from-to)506-511
Number of pages6
JournalCytokine
Volume5
Issue number5
DOIs
Publication statusPublished - 1993 Sep
Externally publishedYes

Keywords

  • Gro/MGSA/recombinant expression/Neutrophil chemoattractant

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Biochemistry
  • Hematology
  • Molecular Biology

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