Detection of the gene encoding programmed cell death-1 (PD-1) in cow genomic DNA isolated form FTA cards
Mulumebet Worku, Ph.D.
Department of Animal Sciences
Programmed cell death 1 (PD-1) encodes an immune checkpoint protein, expressed on white blood cells. Variation in PD-1 is associated with diseases. It is emerging as a target for therapeutics. In cows PD-1 blockade is a potential therapeutic strategy to control viral infection. Thus simple ways of detecting and charactering PD-1 are needed. The objective of this project was to detect the PD-1 gene in cow genomic DNA. Blood was collected from three Holstein Friesian cows on FTA cards. Genomic DNA was extracted following manufactures instructions (Whatman Inc.). The purity and concentration were evaluated using a Nanodrop spectrophotometer. Specific PCR primers for bovine PD-1 were used to detect and amplify the gene. Primers for the house-keeping gene GAPDH were used for controls. The FTA card was easy to collect blood and elute DNA for detection of PD-1. The average concentration of isolated DNA was 20.5 ng/ul and average purity ratio was 1.9. The primers used amplified PD-1 in individual DNA samples ( Average CQ =20.94). The CQ for GAPDH was 20.64. Results will be validated by sequencing. This approach allows for the isolation of high quality DNA of sufficient quantity for detection of PD-1 in cow blood. This simple method for detection of PD-1 gene may be useful in larger studies to assess variation in PD-1 genes and their expression for possible application of targeted immunotherapy in cow health.
Ford, Sade, "Detection of the gene encoding programmed cell death-1 (PD-1) in cow genomic DNA isolated form FTA cards" (2019). Undergraduate Research and Creative Inquiry Symposia. 43.