Detection of Olfactomedin 4 in cow genomic DNA

Student Classification

Senior

Faculty Mentor

Mulumebet Worku, Ph.D.

Department

Department of Animal Sciences

Document Type

Poster

Publication Date

Fall 2019

Disciplines

Animal Sciences

Abstract

Olfactomedin 4 (OLFM4) is an extracellular matrix glycoprotein that belongs to the olfactomedin family. It is involved in cell proliferation, differentiation, apoptosis, adhesion and the inflammatory response to pathogens. Olfactomedin-4 also serves as a pathogenic neutrophil subset marker. Studies are needed to identify and characterize neutrophils subsets in cows and genes involved in the inflammatory response to pathogens. This study aims to detect the OLFM4 gene in bovine genomic DNA. Blood was collected on to FTA cards, from three Holstein Friesian cows, at the NCAT Dairy farm. Genomic DNA was isolated using the manufacturers protocol (Whatman Inc). The purity and concertation of isolated DNA was evaluated using a Nanodrop Spectrophotometer. Specific primers designed for bovine OLFM4 were used to detect the gene with real-time PCR. Primers for the housekeeping gene GAPDH were used as controls. The average concentration of isolated DNA was 67ug/ul and the 260/280 ratio was 1.7. The OLFM4 gene was detected in all three samples tested (Average threshold cycle (Ct) = 29). Olfactomedin-4 might serve as a marker in cow blood. Samples will be sent for sequencing and designed primers will be used to evaluate factors affecting OLFM4 expression on neutrophils subsets in cows and their role inflammatory response to pathogens.

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