Gene-specific universal mammalian sequence-tagged sites: Application to the canine genome
- 61 Downloads
We are developing a genetic map of the dog based partly upon markers contained within known genes. In order to facilitate the development of these markers, we have used polymerase chain reaction (PCR) primers designed to conserved regions of genes that have been sequenced in at least two species. We have refined the method for designing primers to maximize the number that produce successful amplifications across as many mammalian species as possible. We report the development of primer sets for 11 loci in detail:CFTR, COL10A1, CSFIR, CYP1A1, DCN1, FES, GHR, GLB1, PKLR, PVALB, andRB1. We also report an additional 75 primer sets in the appendices. The PCR products were sequenced to show that the primers amplify the expected canine genes. These primer sets thus define a class of gene-specific sequence-tagged sites (STSs). There are a number of uses for these STSs, including the rapid development of various linkage tools and the rapid testing of genomic and cDNA libraries for the presence of their corresponding genes. Six of the eleven gene targets reported in detail have been proposed to serve as “anchored reference loci” for the development of mammalian genetic maps [O'Brien, S. J.,et al., Nat. Genet. 3:103, 1993]. The primer sets should cover a significant portion of the canine genome for the development of a linkage map. In order to determine how useful these primer sets would be for the other genome projects, we tested the 11 primer sets on the DNA from species representing five mammalian orders. Eighty-four percent of the gene-species combinations amplified successfully. We have named these primer sets “universal mammalian sequence-tagged sites” because they should be useful for many mammalian genome projects.
Key wordsgenome mapping evolution homology polymerase chain reaction
Unable to display preview. Download preview PDF.
- Barendse, W., Armitage, S. M., Kossarek, L. M., Shalom, A., Kirkpatrick, B. W., Ryan, A. M., Clayton, D., Li, L., Neibergs, H. L., Zhang, N., Grosse, W. M., Weiss, J., Creighton, P., McCarthy, F., Ron, M., Teale, A. J., Fries, R., McGraw, R. A., Moore, S. S., Georges, M., Soller, M., Womack, J. E., and Hetzel, D. J. S. (1994). A genetic linkage map of the bovine genome.Nat. Genet. 6227.CrossRefPubMedGoogle Scholar
- Bergenhem, N. C. H., Venta, P. J., Hopkins, P. J., and Tashian, R. E. (1992). Variation in coding exons of two electrophoretic alleles at the pigtail macaque carbonic anhydrase I locus as determined by direct, double-stranded sequencing of polymerase chain reaction (PCR) products.Biochem. Genet. 30279.PubMedGoogle Scholar
- ISGN (1987). Guidelines for human gene nomenclature: An international system for human gene nomenclature (ISGN, 1987).Cytogenet. Cell Genet. 4611.Google Scholar
- Li, W.-H., and Grauer, D. (1987).Fundamentals of Molecular Evolution Sinauer Associates, Sunderland, MA.Google Scholar
- Meera Khan, P., Brahe, C., and Wijnen, L. M. M. (1984). Gene map of the dog: Six conserved and three disrupted syntenies.Cytogenet. Cell Genet. 37537.Google Scholar
- Morell, R., Friedman, T. B., Moeljopawiro, S., Hartono, Soewito, and Asher, J. H., Jr. (1992). A frameshift mutation in the HuP2 paired domain of the probable human homolog of murine Pax-3 is responsible for Waardenburg syndrome type 1 in an Indonesian family.Hum. Mol. Genet. 1243.PubMedGoogle Scholar
- Murray, N. E., Brammer, W. J., and Murray, K. (1977). Lambdoid phages that simplify the recovery of in vitro recombinants.Mol. Gen. Genet. 15653.Google Scholar
- Olsen, M., Hood, L., Cantor, C., and Botstein, D. (1989). A common language for physical mapping of the human genome.Science 2451434.Google Scholar
- Sambrook, J., Fritsch, E. F., and Maniatis, T. (1989)Molecular Cloning. A Laboratory Manual (2nd ed.), Cold Springs Harbor Laboratory Press, Cold Springs Harbor, NY.Google Scholar