Overexpression in Escherichia coli and purification of recombinant CI-b1, a Kunitz-type chymotrypsin inhibitor of silkworm

Ningjia He, Hiroshi Fujii, Takahiro Kusakabe, Yoichi Aso, Yutaka Banno, Kohji Yamamoto

研究成果: ジャーナルへの寄稿記事

4 引用 (Scopus)

抄録

Present research provided an efficient approach to obtain large quantities of active recombinant CI-b1, a Kunitz-type chymotrypsin inhibitor of silkworm, Bombyx mori. The cDNA encoding mature CI-b1 was cloned into pDEST17 vector. Recombinant protein with hexa-histidine tag attached to the N-terminal of CI-b1 was expressed in Escherichia coli Origami B cells. It can be purified to homogeneity via the gel filtration chromatography on a Sephacryl S-200 column followed the affinity chromatography on a Ni-NTA column. The two sequential purification procedures yielded 4.3 mg purified (His) 6-tagged CI-b1 from 200 ml of culture medium. Studies on (His) 6-tagged CI-b1 revealed that three disulfide bonds were formed in the recombinant CI-b1 and the inhibitory properties of recombinant CI-b1 against α-chymotrypsin were similar to those of native CI-b1. Recombinant CI-b1 immobilized on Ni-NTA resin was used to detect the interactions occurring between the CI-b1 and its target factors.

元の言語英語
ページ(範囲)9-16
ページ数8
ジャーナルProtein Expression and Purification
38
発行部数1
DOI
出版物ステータス出版済み - 11 1 2004

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His-His-His-His-His-His
Bombyx
Chymotrypsin
Escherichia coli
Affinity Chromatography
Recombinant Proteins
Histidine
Disulfides
Gel Chromatography
Culture Media
B-Lymphocytes
Complementary DNA
Research

All Science Journal Classification (ASJC) codes

  • Biotechnology

これを引用

Overexpression in Escherichia coli and purification of recombinant CI-b1, a Kunitz-type chymotrypsin inhibitor of silkworm. / He, Ningjia; Fujii, Hiroshi; Kusakabe, Takahiro; Aso, Yoichi; Banno, Yutaka; Yamamoto, Kohji.

:: Protein Expression and Purification, 巻 38, 番号 1, 01.11.2004, p. 9-16.

研究成果: ジャーナルへの寄稿記事

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AU - Fujii, Hiroshi

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AU - Aso, Yoichi

AU - Banno, Yutaka

AU - Yamamoto, Kohji

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