Synthesis of a bifunctional metallothionein/beta-glucuronidase fusion protein in transgenic tobacco plants as a means of reducing leaf cadmium levels

Plant J. 1994 Sep;6(3):433-40. doi: 10.1046/j.1365-313x.1994.06030433.x.

Abstract

Chimeric genes under the control of a CaMV 35S promoter with a doubled enhancer (35S2) that encode a mammalian metallothionein (hMTII), or an Escherichia coli beta-glucuronidase (GUS), or a hMTII/GUS fusion protein were introduced into the genome of tobacco (Nicotiana tabacum cv. PBD6). Transcripts and Cd-binding proteins of expected size were observed in plants expressing either the 35S2-hMTII or the 35S2-hMTII/GUS gene, and in the latter plants a protein with GUS activity that was larger than the native GUS enzyme was observed. Thus, plants expressing the hMTII-GUS gene synthesize a bifunctional protein, with both GUS and Cd-binding activity. In an in vitro assay, seedlings expressing either one of these genes had 60-70% lower Cd concentration in their shoots than controls, and Cd translocation to the shoot system was reduced (approximately 20% of Cd absorbed was translocated), compared with that in controls expressing a 35S2-GUS gene, where approximately 50% of the Cd absorbed was translocated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Northern
  • Cadmium / analysis*
  • Glucuronidase / biosynthesis*
  • Metallothionein / biosynthesis*
  • Nicotiana / chemistry
  • Nicotiana / metabolism
  • Plant Leaves / chemistry*
  • Plants, Genetically Modified / chemistry*
  • Plants, Genetically Modified / metabolism
  • Plants, Toxic
  • Recombinant Fusion Proteins / biosynthesis*

Substances

  • Recombinant Fusion Proteins
  • Cadmium
  • Metallothionein
  • Glucuronidase