|Beattie, Craig - UNIVERSITY OF MINNESOTA|
Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: October 7, 1997
Publication Date: N/A
Technical Abstract: To identify bovine epithelial cell genes that are conditionally expressed during pathogen exposure, RNA fingerprinting by arbitrarily primed (RAP)-PCR was used to detect transcripts whose steady-state levels increased when a monolayer cell culture of bovine kidney cells (MDBK) was exposed to E. coli O157:H7 lipopolysaccharide (LPS). Eighty-two combinations of arbitrary 10-mers were used to identify two transcripts that were amplified specifically in tissues exposed to LPS. The intracellular accumulation of these transcripts was verified by ribonuclease protection assay using riboprobes made from cDNA clones. An open reading frame (ORF) deduced from one cDNA clone was homologous to human interleukin 8 (IL-8) whereas an ORF from another cDNA clone was homologous to human growth regulated protein (GRO)-gamma and macrophage inflammatory protein (MIP) 2-beta. IL-8, GRO-gamma, and MIP2-beta are members of the Cys-X-Cys chemokine family that possess chemotactic activity for neutrophils. These results illustrate one system for identifying bovine genes expressed in response to pathogen exposure and provide a means for elucidating the genetic basis for disease susceptibility and resistance.