We developed a genetic mapping resource which allows efficient construction of genetic maps in three-dimensions. In collaboration with leading comparative gene mappers in mice and cattle, we developed 318 PCR-based comparative mapping anchor tagged sequences (CATS) for the construction of Type I (coding gene) comparative genetic maps. This resource can be used to efficiently determine order and distance within any vertebrate species and it allows accurate comparisons across species. The CATS represent coding genes which have been previously mapped by recombination analyses in the mouse and by physical methods in humans. The CATS are evenly spaced at 5 - 10 cM intervals and sequenced PCR products showed that the CATS amplify the correct genes. The primers for the CATS proved to be well conserved across vertebrate species and can be used by any vertebrate mapping project. We developed over 30 collaborations for virtually every species' mapping project throughout the world. The CATS readily detect polymorphism in the feline interspecific backcross pedigree (Felis catus crossed to Prionailurus bengalensis) and have greatly increased the efficiency of the feline genetic map construction. Analyses have shown that approximately 70% of the CATS have variation in our hybrid pedigree detectable by restriction enzymes. Approximately 25-30% of the CATS can be typed by single strand conformation polymorphism detection. Using natural and assisted reproduction, we produced over 61 backcross cats, which is sufficient to develop a 5 cM map of the cat. Linkage of the CATS and the short tandem repeat polymorphisms (STRPs) in the feline interspecific backcross pedigree is currently being analyzed, with mapping emphasis on human chromosomes 11, 12, 4, 6, and X. The feline STRP's proved to be a valuable resource in the genetic map development and are an efficient and reliable resource for genetic individualization in conservation biology and forensic applications. Reciprocal chromosome painting of the feline and human genomes using probes from flow sorted feline and human chromosomes increased our knowledge of comparative genome organization for these two species. This technique supported our data that the feline genome is organizationally well conserved to the human genome, in comparison to other species' genomes and this data provided new evidence for the reconstruction of the mammalian radiation.