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Type of Document Dissertation Author McNally, Lacey R. URN etd-04122004-171939 Title Chromosomal Localization of a Proinsulin Transgene Inserted with a Transposon-Based Vector into Japanese Quail, Coturnix Coturnix Degree Doctor of Philosophy (Ph.D.) Department Veterinary Microbiology & Parasitology (Veterinary Medical Sciences) Advisory Committee
Advisor Name Title Richard Cooper Committee Chair Fred Enright Committee Member William Henk Committee Member William Todd Committee Member Gale Bateman Dean's Representative Keywords
- primed in situ hybridization
- laser microdissection
- transgene
- Japanese quail
Date of Defense 2004-04-02 Availability unrestricted Abstract The overall goals of this research were to develop a reproducible method of detecting stable DNA insertion into Japanese quail and provide a method for gene location on avian chromosomes. This research resulted in the development of a different method of obtaining chromosome spreads in Japanese quail, the establishment of primed in situ hybridization as a method for the chromosomal gene detection in birds, development of Teflon-coated coverslip slides to facilitate laser microdissection of 0.5 ƒİm samples, and chromosomal identification of proinsulin transgene insertions by laser microdissection and nucleotide sequence from G2 Japanese quail. The 28S rDNA was found on a macrochromosome and a microchromosome pair by primed in situ hybridization, fluorescent in situ hybridization, and silver staining. Teflon-coated coverslip slides were created to facilitate laser microdissection of avian chromosomes for DNA amplification and nucleotide sequencing. Transgenic G2 Japanese quail produced in Dr. Richard Cooper¡¦s laboratory were identified by laser microdissection and found to have 2-5 chromosomal insertions of the proinsulin transgene.
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