This collaborative research project is designed to provide a physical and genetic map of the genome of Drosophila Melanogaster, using in situ hybridization of large DNA molecules cloned into yeast cells by means of yeast artificial chromosome (YAC) vectors. Preliminary experiments have yielded an initial YAC library containing Drosophila inserts of up to 140 kbp, some of which are presently being studied, and construction of a library with even larger inserts is in progress. Library construction has been carried out using a new YAC vector containing the ends of the transposable element P with an eye to P-element-mediated transformation of large molecules of DNA.
The specific aims of the proposed research are to produce and characterize a Drosophila library with inserts averaging 165 kbp (ideally using a vector under construction which will allow easy production of RNA probes for in situ hybridization), to perform initial tests of the feasibility of transformation using YAC vectors, and (the main thrust of the proposed research) to determine the location of large-insert YAC clones in the Drosophila genome by means in situ hybridization to salivary gland chromosomes.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
8R01HG000357-03
Application #
3333508
Study Section
Special Emphasis Panel (SRC)
Project Start
1988-09-30
Project End
1993-08-31
Budget Start
1990-09-01
Budget End
1991-08-31
Support Year
3
Fiscal Year
1990
Total Cost
Indirect Cost
Name
Washington University
Department
Type
Schools of Medicine
DUNS #
062761671
City
Saint Louis
State
MO
Country
United States
Zip Code
63130
Hartl, D L; Ochman, H (1996) Inverse polymerase chain reaction. Methods Mol Biol 58:293-301
Russell, R J; Robin, G C; Kostakos, P et al. (1996) Molecular cloning of an alpha-esterase gene cluster on chromosome 3r of Drosophila melanogaster. Insect Biochem Mol Biol 26:235-47
Hartl, D L; Ochman, H (1994) Inverse polymerase chain reaction. Methods Mol Biol 31:187-96
Lozovskaya, E R; Petrov, D A; Hartl, D L (1993) A combined molecular and cytogenetic approach to genome evolution in Drosophila using large-fragment DNA cloning. Chromosoma 102:253-66
Hartl, D L; Lozovskaya, E R (1992) The Drosophila genome project: current status of the physical map. Comp Biochem Physiol B 103:1-8
Carulli, J P; Hartl, D L (1992) Variable rates of evolution among Drosophila opsin genes. Genetics 132:193-204
Hartl, D L; Ajioka, J W; Cai, H et al. (1992) Towards a Drosophila genome map. Trends Genet 8:70-5
Palazzolo, M J; Sawyer, S A; Martin, C H et al. (1991) Optimized strategies for sequence-tagged-site selection in genome mapping. Proc Natl Acad Sci U S A 88:8034-8