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Genotypic and Phenotypic Characterization of Penicillium marneffei Mutants Produced by Agrobacterium-Mediated Transformation

Price, Eric C.

Abstract Details

2012, Master of Science in Biological Sciences, Youngstown State University, Department of Biological Sciences and Chemistry.

The dimorphic fungus Penicillium marneffei is an etiological agent of penicillosis and the third leading cause of AIDS related death in Southeast Asia. This fungus is unique to Penicillium species in that it displays thermal dimorphism. Specifically, at 25°C P. marneffei grows as a filamentous mold, whereas at 37°C hyphae or conidia differentiate into fission yeast.

One of the key factors to this organism’s pathogenicity is the dimorphic switch that it performs in vivo to establish an infection. Mutagenesis with focus on morphology has been used to discover certain pathways linked to dimorphism in P. marneffei. The technique of Agrobacterium-mediated transformation was used to produce random T-DNA insertion mutants. From this prior work, two mutants with outward phenotypic defects were chosen to be characterized. Initially, the mutants were characterized phenotypically by slide culture techniques to note any differences in morphology as compared to the wild type at 25°C and 37°C. The disrupted genes of each mutant were identified by amplification of the flanking regions of the T-DNA insertion through inverse PCR and sequencing. These results indicated that mutant I209 possessed a defect in the septin gene, designated aspC, whereas the mutation in the second mutant, strain I219, was found to be in the gene encoding 3-methylcrotonyl CoA carboxylase (MccB). The expression of the interrupted gene in mutant I209 was verified using RT-PCR. Verification of a single T-DNA insertion was confirmed by Southern blot analysis.

Chester R. Cooper Jr., PhD (Advisor)
David K. Asch, PhD (Committee Member)
Jonathan Caguiat, PhD (Committee Member)
64 p.

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Citations

  • Price, E. C. (2012). Genotypic and Phenotypic Characterization of Penicillium marneffei Mutants Produced by Agrobacterium-Mediated Transformation [Master's thesis, Youngstown State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1341255805

    APA Style (7th edition)

  • Price, Eric. Genotypic and Phenotypic Characterization of Penicillium marneffei Mutants Produced by Agrobacterium-Mediated Transformation. 2012. Youngstown State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=ysu1341255805.

    MLA Style (8th edition)

  • Price, Eric. "Genotypic and Phenotypic Characterization of Penicillium marneffei Mutants Produced by Agrobacterium-Mediated Transformation." Master's thesis, Youngstown State University, 2012. http://rave.ohiolink.edu/etdc/view?acc_num=ysu1341255805

    Chicago Manual of Style (17th edition)