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United States Department of Agriculture

Agricultural Research Service

Project Overview


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"We strive to discover novel genetic tools to improve the
agronomic properties of potatoes and tree fruits."


Project Overview

 Harvesting Potatoes Specific Research Strategies
  • Mobilizing transgenes into potato cultivars to confer resistance to microbial pathogens.
  • Continuous investigation and characterization of agronomically useful genes from wild Solanum species.
  • Application of genomics to citrus improvement.
  • Down-regulation of undesirable glycoalkaloid toxicants and  tuber bruise reactions using antisense or siRNA technology.
 Bill and Paul Characterization & Distribution of Genetic Resources

We continue to clone and characterize agronomically useful control elements and vectors for use in the research community.

 Gene Map Characterization & Mapping of Repetitive DNA Elements

We seek to discover important mechanisms involved in gene evolution and function by mapping and characterizing repetitive DNA elements within the genome.


Vectors & Control Elements

We make these plasmid maps of the vectors and control elements used in our lab available for viewing and downloading. To view them, you must download and install Adobe Reader® from Adobe Systems, Inc. Please contact Dr. Bill Belknap for sequences and maps.

Control Elements

Granule Bound Starch Synthase (GBSS) promoter
View Map(PDF)
  • HindII-EcoRI fragment in pCR vector
  • High-level general tuber and root expression
  • Ref: Van der Lief, et. al., Mol. Gen. Genet., 228:240-248, 1991.
Ubiquitin 3 Promoter
View Map(PDF)
  • GUS expression cassette in pUC 19 vector
  • Constitutive expression
  • Ref: Garbarino et. al., Plant Physiology 109: 1371-1378, 1995.
Ubiquitin 3 poly ubiquitin fusion promoter
View Map(PDF)
  • GUS expression cassette in pUC 19 vector
  • High-level constitutive protein expression
  • Ref: Garbarino et. al., Plant Physiology 109: 1371-1378, 1995.
Ubiquitin 3 Terminator
View Map(PDF)
  • EcoRI insert in pCR vector
  • All-purpose polyadenlyation signal
Patatin Class I Promoter in pUC 19 vector
View Map(PDF)
  • Tuber specific expression
  • Ref: Blundy, et. al., Plant Molecular Biology, 16:153-160, 1991.
Granule Bound Starch Synthase (GBSS) promoter + Ubiquitin3 terminator
View Map(PDF)
  • HindIII-EcoRI expression cassette with BamHI cloning site for gene insertion
  • Expression in tubers and roots
409s-MCS-409T Detail
View Map(PDF)

409s-MCS-409T in pBinplus/ARS
View Map(PDF)

Binary Vectors

pBINPLUS-ARS
View Map (PDF)
  • Binary vector developed from pBINPLUS
  • Complete sequence available in GenBank accession DQ320121
ubi3-GUS-ubi3T
View Map(PDF)
  • in pBINPLUS/ARS

Laboratory Protocols

We make these experimental protocols used in our lab available for viewing and downloading. To view them, you must download and install Adobe Reader® from Adobe Systems, Inc. Please contact Dr. Bill Belknap for more information.

  • [ PDF] Agrobacterium-mediated Transformation of Potato Stems
  • [ PDF] Sterile Propagation of Potato Plantlets and Microtubers
  • [ PDF] Electrocompetent Agrobacterium
  • [ PDF] Benedict's Test for Agrobacterium
  • [ PDF] Preparation of Potato Tubers for RNA Extraction
  • [ PDF] Miniprep for RNA
  • [ PDF] RNA Extraction from Potato Leaf
  • [ PDF] RNA Extraction from Potato Tubers

Photo Gallery

 

Tuber Initiation

Tuber Initiation

Potato tubers (cv. lenape) beginning to emerge from nodal cuttings on tuberization media
Tubers Ready for Transformation

Tubers ready for transformation

Potato tubers (cv. lenape) at proper size for Agrobacterium infection
Stage 3 Selection

Stage 3 Selection

Transgenic shoot rooting on Stage 3 media with 200mg/L Kanamycin
Planting into Vermiculite

Planting into vermiculite

Transfer of plantlet from gel media to vermiculite
Positive Benedicts Test

Positive Benedicts test for Agrobacterium


Last Modified: 5/10/2011