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. 1997 Nov;25(1-3):235–238. doi: 10.1023/A:1007976103088

Preparation of recombinant six-histidine-tagged human LECT2, a chemotactic protein to neutrophils, in Escherichia coli

Mie Ito, Satoshi Yamagoe, Kazuo Tomizawa, Satoshi Mizuno, Masaru Tanokura, Kazuo Suzuki
PMCID: PMC3466745  PMID: 22358898

Abstract

LECT2 is a chemotactic protein to neutrophils. A recombinant six-histidine-tagged human LECT2, (His)6-LECT2, was expressed in E. coli using a pET21a(+) vector. The (His)6-LECT2 was purified from the soluble fraction in E. coli as a single band in sodium dodesyl sulfate/polyacrylamide gel electrophoresis using three steps of column chromatography with Ni2+-charged nitrilo-triacetic acid (Ni-NTA) agarose, DEAE-Sepharose, and CM-Sepharose. The purified (His)6-LECT2 was yielded with 96 µg from the soluble fraction of 1,500 ml culture of E. coli. The circular dichroism spectrum of (His)6-LECT2 showed the folded structure, which is rich in β-sheet structure and rare in α-helix.

Keywords: cytokine, LECT2, human, recombinant, six-histidine-tag, E. coli

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