Comparative fusion expression of maize SINAT5 in two different strains of Escherichia coli.

Genet Mol Res

Department of Molecular Biotechnology, Research Institute for Fundamental Sciences, University of Tabriz, Tabriz, Iran.

Published: August 2012

AI Article Synopsis

  • SINAT5 is a plant E3 ligase that influences auxin signaling and root development by targeting the NAC1 protein for ubiquitination.
  • Efficient production and purification of this protein are crucial for further studies on its functional roles.
  • The study produced a "maltose binding protein-sinat5" fusion in two E. coli strains, yielding 33 mg of protein per liter in TB1 cells and 18 mg in DH5α cells, with consistent expression patterns observed in both cultures.

Article Abstract

SINAT5 is a plant E3 ligase that regulates auxin signaling and root morphogenesis by ubiquitination of the NAC1 protein. Consequently, it may be a putative regulator of aspects of plant development cycles that are controlled by auxin. Efficient production, purification and correctly folded form of this protein are important requirements for functional studies. We produced and quantitatively compared fusion expression of the "maltose binding protein (mbp)-maize sinat5" construct in two different strains of Escherichia coli. One-step purification of fused products gave about 33 mg protein/L bacterial cell culture for E. coli TB1 cells and approximately 18 mg protein/L bacterial cell culture for E. coli DH5α cells. Continuous expression of the fused product and similarity of growth patterns were observed in both cultures.

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http://dx.doi.org/10.4238/2012.August.24.1DOI Listing

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