1887

Abstract

The enhanced green fluorescent protein (EGFP) gene is a modified version of the green fluorescent protein gene of the jellyfish with a codon usage that corresponds well to that found in many GC-rich streptomycete genes. Here the use of EGFP as a reporter for the analysis of spatially and temporally regulated gene expression in A3(2) is demonstrated. The EGFP gene was inserted into plasmids that can replicate in , greatly facilitating the construction of EGFP gene fusions. The plasmids can be transferred readily to by conjugation, whereupon two of them (pIJ8630 and pIJ8660) integrate at the chromosomal attachment site for the temperate phage ϕC31. These vectors were used to analyse the spatial and temporal expression of , which encodes a σ factor required for spore maturation, and of , a pathway-specific regulatory gene for the production of undecylprodigiosin, one of the four antibiotics made by . While transcription of appeared to be confined to developing and mature spore chains, transcription of occurred only in ageing substrate mycelium. A further plasmid derivative (pIJ8668) was made that lacks the ϕC31 attachment site, allowing the EGFP gene to be fused transcriptionally to genes of interest at their native chromosomal locations.

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1999-09-01
2019-10-18
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