2007 NSTI Nanotechnology Conference and Trade Show - Nanotech 2007 - 10th Annual

Galactosidase Gene Expression in Yeast Cells Enhanced by Nanoparticles

S-F Chien, S-H Chen and C-T Lin
Northern Illinois University, US

Keywords:
gene expression, yeast cell, nanoparticle, therapeutic protein

Abstract:
We report the enhancement of gene expression for rice -galactosidase gene ( -Gal) in yeast cells assisted by metal nanomaterials. Au and Ag nanoparticles, and their nanocomposites, silica@Au and silica@Ag were prepared and characterized by UV-vis spectroscopy and SEM technique. We have cloned rice -galactosidase gene into the yeast chromosome, where the cloned cells were pre-cultured and induced into a medium containing each of the testing nanomaterials. The enzyme activity was determined by a synthetic substrate, p-nitrophenyl- -D-galctopyranoside, and the yellow product yield was read in a spectrophotometer at 400 nm. When Au and Ag nanoparticles were added to the culture, a 3-5 fold enhancement in -galactosidase was observed for intracellular activity as well as the secreted activity into the medium. The secreted protein was analyzed to have a pure form and displayed as a single protein band in the SDS-gel electrophoresis. The plausible mechanism on the effect of nanomaterials in gene expression for the rice -galactosidase gene in yeast cells will be discussed.

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