| study was undertaken to genetically manipulate a cyanobacterium to generate a recombinant organism capable of increased metal sequestration. The work represents one of the first reports of expression of a non-photosynthetic eukaryotic gene in cyanobacteria.;A cloned copy of the Saccharomyces cerevisiae copper metallothionein gene CUPI, responsible for enhanced metal sequestration in yeast, was employed in a vector construct developed for a naturally competent, unicellular cyanobacterium. For utilization in PCR amplification, forward and reverse primers containing restriction endonuclease sequences were designed to allow directional cloning into the cyanobacterial shuttle vector pTrcIK. The recombinant plasmid was incorporated via homologous recombination into the genome of R2-PIM8 ;The cyanobacterial vector construct carrying the cloned CUPI gene, pMcK2, was shown to be capable of directing in vitro inducible protein expression as determined by studies utilizing... |
