Expression of the Copper Metallothionein CUPI from Saccharomyces cerevisiae in the Cyanobacterium Synechococcus R2-PIM8(smtA)

Authors

Patricia M. McCormick, University of Southern Mississippi
Gordon C. Cannon, University of Southern MississippiFollow
Sabine Heinhorst, University of Southern MississippiFollow

Document Type

Article

Publication Date

3-1-1999

Department

Chemistry and Biochemistry

School

Mathematics and Natural Sciences

Abstract

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 ¹⁰⁹Cd²⁺ 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).

Publication Title

Current Microbiology

Volume

38

Issue

3

First Page

155

Last Page

162

Recommended Citation

McCormick, P. M., Cannon, G. C., Heinhorst, S. (1999). Expression of the Copper Metallothionein CUPI from Saccharomyces cerevisiae in the Cyanobacterium Synechococcus R2-PIM8(smtA). Current Microbiology, 38(3), 155-162.
Available at: https://aquila.usm.edu/fac_pubs/9221