Expression of human transforming growth factor β1 in E. coli (I) - Direct expression in cytoplasm

Jianyin Long; Huixin Wang; Li Liu; Fang Wang; Tingchong Zhou

doi:10.1007/BF02882894

Expression of human transforming growth factor β1 in E. coli (I) - Direct expression in cytoplasm

Jianyin Long, Huixin Wang, Li Liu, Fang Wang, Tingchong Zhou

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

Abstract

PCR technique is used to amplify the mature peptide gene of human transforming growth factor β1 (hTGFβ1); the gene is verified by full-length sequence analysis. In DH5α/pBV220 expression system, hTGFβ1 attains expression in the cytoplasm of E. coli up to 16% . The recombinant protein is proved to be the monomer of hTGFβ1 by N-terminal amino acids analysis and immunoblotting. After refolding of the monomer protein in vitro in glutathione system or CHPAS/DMSO system, the dimeric protein accumulates to 30% in the refolding mixture. The recombinant protein is purified to homogeneity on silver staining, and is shown to have strong biological activity from MTT bioassay on Mv1Lu cells.

Original language	English (US)
Pages (from-to)	548-554
Number of pages	7
Journal	Science in China, Series C: Life Sciences
Volume	41
Issue number	5
DOIs	https://doi.org/10.1007/BF02882894
State	Published - 1998
Externally published	Yes

Keywords

Assembly and renaturation in vitro
E. Coli
Expression in cytoplasm
Tgfβ1

ASJC Scopus subject areas

General Biochemistry, Genetics and Molecular Biology
General Environmental Science
General Agricultural and Biological Sciences

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10.1007/BF02882894

Cite this

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title = "Expression of human transforming growth factor β1 in E. coli (I) - Direct expression in cytoplasm",

abstract = "PCR technique is used to amplify the mature peptide gene of human transforming growth factor β1 (hTGFβ1); the gene is verified by full-length sequence analysis. In DH5α/pBV220 expression system, hTGFβ1 attains expression in the cytoplasm of E. coli up to 16% . The recombinant protein is proved to be the monomer of hTGFβ1 by N-terminal amino acids analysis and immunoblotting. After refolding of the monomer protein in vitro in glutathione system or CHPAS/DMSO system, the dimeric protein accumulates to 30% in the refolding mixture. The recombinant protein is purified to homogeneity on silver staining, and is shown to have strong biological activity from MTT bioassay on Mv1Lu cells.",

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author = "Jianyin Long and Huixin Wang and Li Liu and Fang Wang and Tingchong Zhou",

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journal = "Science in China, Series C: Life Sciences",

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TY - JOUR

T1 - Expression of human transforming growth factor β1 in E. coli (I) - Direct expression in cytoplasm

AU - Long, Jianyin

AU - Wang, Huixin

AU - Liu, Li

AU - Wang, Fang

AU - Zhou, Tingchong

PY - 1998

Y1 - 1998

N2 - PCR technique is used to amplify the mature peptide gene of human transforming growth factor β1 (hTGFβ1); the gene is verified by full-length sequence analysis. In DH5α/pBV220 expression system, hTGFβ1 attains expression in the cytoplasm of E. coli up to 16% . The recombinant protein is proved to be the monomer of hTGFβ1 by N-terminal amino acids analysis and immunoblotting. After refolding of the monomer protein in vitro in glutathione system or CHPAS/DMSO system, the dimeric protein accumulates to 30% in the refolding mixture. The recombinant protein is purified to homogeneity on silver staining, and is shown to have strong biological activity from MTT bioassay on Mv1Lu cells.

AB - PCR technique is used to amplify the mature peptide gene of human transforming growth factor β1 (hTGFβ1); the gene is verified by full-length sequence analysis. In DH5α/pBV220 expression system, hTGFβ1 attains expression in the cytoplasm of E. coli up to 16% . The recombinant protein is proved to be the monomer of hTGFβ1 by N-terminal amino acids analysis and immunoblotting. After refolding of the monomer protein in vitro in glutathione system or CHPAS/DMSO system, the dimeric protein accumulates to 30% in the refolding mixture. The recombinant protein is purified to homogeneity on silver staining, and is shown to have strong biological activity from MTT bioassay on Mv1Lu cells.

KW - Assembly and renaturation in vitro

KW - E. Coli

KW - Expression in cytoplasm

KW - Tgfβ1

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JO - Science in China, Series C: Life Sciences

JF - Science in China, Series C: Life Sciences

IS - 5

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Expression of human transforming growth factor β1 in E. coli (I) - Direct expression in cytoplasm

Abstract

Keywords

ASJC Scopus subject areas

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