Title page for ETD etd-11212012-040023

Type of Document Master's Thesis
Author Comeaux, Jay Louis
URN etd-11212012-040023
Title Transfer of plasmids by genetically-engineered Erwinia carotovora
Degree Master of Science
Department Biology
Advisory Committee
Advisor Name Title
Cairns, John Jr. Committee Chair
Lacy, George H. Committee Member
Wallace, Bruce Committee Member
  • Molecular biology
Date of Defense 1989-12-05
Availability restricted

The ability of a genetically-engineered Erwirzia carotovora subsp. carotovora (Ecc) strain to transfer recombinant chromosomal DNA or plasmids to wildtype Ecc or Pseudomonas fluorescens was tested on filters, within soil microcosms, and in planta. Ecc was engineered by chromosomal insertion of a disarmed endo-pectate lyase gene marked with a 1.4kb DNA fragment conferring kanamycin resistance. Plasmids RPI and pBR322 were introduced separately into engineered Ecc clones. These strains served as donors in genetic transfer experiments. No transfer of the inserted kan marker or of pBR322 was observed under any experimental condition. In filter matings, RPI was transferred to wildtype Ecc at a frequency of 3.6 X 10-2 transconjugants per donor (TPD) and to P. fluorescens at a frequency of 2.4 X 10-5 TPD. In matings conducted in potato tubers inoculated using sewing needles, the respective frequencies were 4.0 X 10-4 and 2.0 X 10-4, while matings on potato slices yielded frequencies of 4.7 X 10-2 and 2.3 X 10-2. In soil microcosms, the maximum transfer frequencies observed were 2.3 X 10-3 and 8.4 X 10-5 TPD.

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