Gene Cloning, Bacterial Expression, and Purification of a Novel Rice (Oryza sativa L.) Ubiquitin-Related Protein, RURM1

Takuji TSUKIYAMA; Jongwon LEE; Yutaka OKUMOTO; Masayoshi TERAISHI; Takatoshi TANISAKA; Kuniyo INOUYE

doi:10.1271/bbb.90692

Biochemistry & Molecular Biology Notes

Gene Cloning, Bacterial Expression, and Purification of a Novel Rice (Oryza sativa L.) Ubiquitin-Related Protein, RURM1

Takuji TSUKIYAMA, Jongwon LEE, Yutaka OKUMOTO, Masayoshi TERAISHI, Takatoshi TANISAKA, Kuniyo INOUYE

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Keywords: Escherichia coli, glutathione S-transferase (GST)-fusion protein, Oryza sativa, recombinant protein, ubiquitin-like protein

JOURNAL FREE ACCESS

2010 Volume 74 Issue 2 Pages 430-432

DOI https://doi.org/10.1271/bbb.90692

View "Advance Publication" version (February 7, 2010).

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Abstract

A rice ubiquitin-related modifier-1 (Rurm1) gene was cloned and transformed in Escherichia coli. We successfully expressed the RURM1 protein as a glutathione S-transferase (GST)-fusion protein by cultivating the E. coli cells at 16 °C for 16 h. After cleavage of GST, we obtained a single protein of 12 kDa. This protein was identified as the RURM1 protein by western blot analysis.

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