A PCR method employing single arbitrarily chosen primers (Arbitrarily primed PCR) generates consistent 'fingerprints' for the mouse genome. These fingerprints include DNA fragment polymorphisms which can be genetically mapped in recombinant inbred lines of mice. This method is very fast and simple and uses only a few nanograms of template DNA. Primers can be used individually or in almost every pairwise combination. There are an average of over four mappable polymorphisms unique to each primer or pair of primers in the BXD recombinant inbred lines. We will be able to use as few as 35 PCR primers to place an additional 2400 DNA polymorphisms on the mouse genetic map in three years using the 26 BXD, 25 AXB and 25 BXA recombinant inbred lines. The accuracy of this map should be better than 5 centiMorgans and should have an average of almost one polymorphism per million base pairs. The genetically assigned AP-PCR fragments will be a valuable resource along with the many restriction fragment length polymorphisms (RFLPs) and PCR based (CA)n polymorphisms already on the map. Those polymorphisms that co-segregate with genetically mapped mutations should be useful for cloning the regions around the genes responsible for such mutations. For instance, the BXD, AXB and BXA recombinant inbred lines are models for susceptibility to certain diseases such as atherosclerosis, glucocorticoid induced cleft palate, some infections and some neoplasms. Mapped AP-PCR polymorphisms will also facilitate the genetic localization of mouse genomic clones that are being assembled into a contig map. Furthermore, due to synteny, information about the mouse genome can sometimes be extended to humans and vice versa.

Agency
National Institute of Health (NIH)
Institute
National Human Genome Research Institute (NHGRI)
Type
Research Project (R01)
Project #
5R01HG000456-02
Application #
2208841
Study Section
Genome Study Section (GNM)
Project Start
1992-03-15
Project End
1995-02-28
Budget Start
1993-03-01
Budget End
1994-02-28
Support Year
2
Fiscal Year
1993
Total Cost
Indirect Cost
Name
California Institute of Biological Research
Department
Type
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92037
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Ralph, D; Postic, D; Baranton, G et al. (1993) Species of Borrelia distinguished by restriction site polymorphisms in 16S rRNA genes. FEMS Microbiol Lett 111:239-43
al-Janabi, S M; Honeycutt, R J; McClelland, M et al. (1993) A genetic linkage map of Saccharum spontaneum L. 'SES 208'. Genetics 134:1249-60
Ralph, D; Que, Q; Van Etten, J L et al. (1993) Leptospira genomes are modified at 5'-GTAC. J Bacteriol 175:3913-5
Welsh, J; Chada, K; Dalal, S S et al. (1992) Arbitrarily primed PCR fingerprinting of RNA. Nucleic Acids Res 20:4965-70