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  • Copper induction of enhanced green fluorescent protein expression in Pleurotus ostreatus driven by laccase poxa1b promoter.

Copper induction of enhanced green fluorescent protein expression in Pleurotus ostreatus driven by laccase poxa1b promoter.

FEMS microbiology letters (2012-10-12)
Antonella Amore, Yoichi Honda, Vincenza Faraco
ABSTRACT

In silico analyses of several laccase promoter sequences have shown the presence of many different responsive elements differentially distributed along the promoter sequences. Analysis of Pleurotus ostreatus laccase promoter poxa1b extending around 1400-bp upstream of the start codon showed the presence of several putative response elements, such as 10 metal-responsive elements. Development of a system for in vivo analysis of P. ostreatus laccase promoter poxa1b by enhanced green fluorescent protein expression was carried out, based on a polyethylene glycol-mediated procedure for fungal transformation. Quantitative measurement of fluorescence expressed in P. ostreatus transformants grown in the presence and in the absence of copper sulfate was performed, demonstrating an increase in expression level induced by the metal.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Copper(II) sulfate, JIS first grade, ≥97.5%
Sigma-Aldrich
Copper(II) sulfate, anhydrous, powder, ≥99.99% trace metals basis
Sigma-Aldrich
Copper(II) sulfate solution, 4 % (w/v) (prepared from copper (II) sulfate pentahydrate)
Sigma-Aldrich
Fehling′s reagent (A)
Sigma-Aldrich
Copper(II) sulfate, puriss. p.a., anhydrous, ≥99.0% (RT)
Sigma-Aldrich
Copper(II) sulfate, ReagentPlus®, ≥99%
Sigma-Aldrich
Copper(II) sulfate, puriss., meets analytical specification of Ph. Eur., BP, USP, anhydrous, 99-100.5% (based on anhydrous substance)