SURVEY OF PRION GENE SEQUENCES OF WILD ANIMALS IN BLACK HILLS

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This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. This primary goal of this project is to identify the sequences of the prion protein gene (PRNP) in various animals. The two main areas of interest are the evolutionary history of the gene and possible serial transmission. The main focus thus of this project is to sequence PRNP from carnivores and other animals common to the Black Hills region. The reason carnivores were chosen is because of the real possibility of them consuming the flesh of animals that have been documented to contract this progressive, fatal disease. We have obtained existing tissue samples from area trappers, law enforcement officers and taxidermists, and have extracted DNA from the following animals; coyote, badger, raccoon, bobcat, lynx, skunk and pronghorn antelope. The PRNP coding region has been sequenced from each of these animals using degenerate primer sets to obtain partial coding sequence. New species specific primers will be designed in order to fully sequence the coding regions of each of these animals. Additionally, an attempt will be made to obtain sequence data for certain areas of the promoter region of the PRNP gene. All sequence data will be compared to a national database of previously sequenced animals in hopes of identifying similarities that would give some insight to the evolutionary history of the gene as well as the possibility of serial transmission.