Title page for ETD etd-04122004-171939


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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