Applied Microbiology and Biotechnology, Vol.52, No.6, 806-810, 1999
Metal-induced expressing of mammal Metallothionein-1 gene in cyanobacteria to promote cadmium-binding preferences
The metal(zinc)-inducible smtA gene promoter (smt O-P) from cyanobacteria was applied for the expression of mouse MT-1 cDNA in the filamentous cyanobacterium Anabaena sp. PCC 7120 to enhance its metal-binding capability and to change its main binding specificity from zinc to cadmium. Shuttle expression vector pKT-MRE transformed the cyanobacterial cells by triparent conjugal transfer. Positive clones were screened and identified by streptomycin, DNA dot blot, SDS-PAGE and Western blot analysis. Photosynthetic oxygen evolution and metal atom absorption indicated that under the cadmium stress the metal-induced expression of foreign mMT-1 doubled their cadmium resistance and developed cells showing a much higher preference to absorb cadmium other than zinc in medium. The cadmium content in cell extract rose from 11% to 36%, and the cadmium cleared from media by transgenic cells rose from 18% to 62%. There was only a slight enhancement for zinc binding in the wild or transgenic type.
Keywords:PROKARYOTIC METALLOTHIONEIN;LOCUS