SURVEY OF PRION GENE SEQUENCES OF WILD ANIMALS IN BLACK HILLS

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。这个项目的主要目标是鉴定各种动物的朊蛋白基因（PRNP）序列。两个主要领域的兴趣是基因的进化史和可能的系列传播。因此，该项目的主要重点是对布莱克山地区常见的食肉动物和其他动物的PRNP进行测序。选择食肉动物的原因是因为它们确实有可能食用动物的肉，而这些动物已经被记录在案，会感染这种渐进的、致命的疾病。我们从地区猎人、执法人员和标本剥制师那里获得了现有的组织样本，并从以下动物身上提取了DNA；郊狼、獾、浣熊、山猫、猞猁、臭鼬和叉角羚。利用退化引物集对这些动物的PRNP编码区进行了测序，获得了部分编码序列。新的物种特异性引物将被设计出来，以便对每种动物的编码区域进行完整的测序。此外，将尝试获得PRNP基因启动子区域某些区域的序列数据。所有序列数据将与先前测序的动物的国家数据库进行比较，希望找出相似性，从而对基因的进化史以及序列传播的可能性有所了解。

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.