Expression of the copper metallothionein CUPI from Saccharomyces cerevisiae in the cyanobacterium Synechococcus R2-PIM8(smtA)

PM McCormick
Gordon C. Cannon, University of Southern Mississippi
Sabine Heinhorst, University of Southern Mississippi


The coding sequence for Saccharomyces cerevisiae copper metallothionein (CUPI), the protein responsible for enhanced sequestration of Cu2+ in yeast, was placed under the control of an inducible synthetic Escherichia coli promoter in the cyanobacterial vector pTrcIK. Strain R2-PIM8(smtA) of Synechococcus sp. PCC 7942 was transformed with the resulting construct pMcK2, and the yeast CUPI gene was integrated into its genome via homologous recombination. The pMcK2 plasmid directed the synthesis of a protein product of the expected size in an in vitro E. coli transcription/translation system. In the transgenic cyanobacteria, the integrated CUPI gene was transcribed and produced a protein product with the expected metallothionein characteristics, as determined by Cd-109(2+) binding assays. At this level of expression, the yeast metallothionein, although functional, did not increase the tolerance range of the transgenic Synechococcus to Cu2+ Or Cd2+ beyond that of the untransformed R2-PIM8(smtA).