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SfMTP1 and SpMTP1 cloning and expression in Saccharomyces cerevisiae: Potential for the genetic engineering of Arabidopsis thaliana

Posted on:2014-10-03Degree:M.SType:Thesis
University:The University of Texas - Pan AmericanCandidate:Garza, Elyssa RaeFull Text:PDF
GTID:2450390005488967Subject:Biology
Abstract/Summary:
To evaluate metal tolerance conferred by the overexpression of Metal Tolerance Protein 1 (MTP1) in transgenic non-accumulator plants such as Arabidopsis thaliana, a model system was developed in Saccharomyces cerevisiae containing either SfMTP1 cDNA, from the metal tolerant plant Streptanthus farnsworthianus, or SpMTP1 cDNA, from the hyperaccumulator Streptanthus polygaloides. Nonquantitative RT-PCR showed constitutive expression of MTP1 in both plant species. Alignment of SfMTP1 and SpMTP1 protein sequences shows a gap in SpMTP1 in a variable histidine region known to affect metal transport. Disk assays revealed SfMTP1 cDNA imparted greater Ni tolerance to yeast than SpMTP1 cDNA; however, SpMTP1 showed higher Cd tolerance than SfMTP1. No significant difference was found with Co; but both cDNAs made yeast slightly more Zn sensitive. MTP1 histidine-rich regions control metal transport mechanisms, but differences in these regions may be responsible for higher tolerance of specific metals between different Streptanthus species.
Keywords/Search Tags:MTP1, Metal, Tolerance, Spmtp1, Sfmtp1
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