We found a match
Your institution may have access to this item. Find your institution then sign in to continue.
- Title
Using microarray analysis to decipher gene expression in mastitis causing Escherichia coli exposed to bovine whey.
- Authors
Worku, M.; Bowman-Simpson, J.; Matterson, P.
- Abstract
Escherichia coli cause mastitis upon entry into the mammary gland where exposure to milk components such as whey, occurs. Infection with Escherichia coli, release of endotoxin and the resulting inflammation is associated with changes in milk composition, loss of production and has consequences for animal health. Exposure to immune factors in whey are important in combating infection through immunomodualtion and can impact bacterial pathogenesis. The objective of this study was to evaluate the effect of host immune factors in whey on gene expression in E.coli using microarray analysis. A mid-log culture of E.coli isolated from an acute case of clinical mastitis was grown. Whey samples were prepared from clinically healthy cows. The samples were heat inactivated (56°C, 30 min.). Six samples of E. coli (109) were incubated in RNase free Phosphate Buffered Saline (PBS) as negative controls. Six samples were incubated with a 1:1 dilution of the inactivated whey (10 min., 37°C). RNA from control and treated samples was isolated using the RNeasy (Quaigen) kits. The integrity and size distribution of total purified RNA was checked on ethidium bromide stained denaturing agarose gels and using a bioanlyzer. Two E. coli K12 Starter V2 array chips consisting of 2 identical grids with a total of 192 spots (MWG Biotech, High Point) were used for expression profiling of E. coli. Data were analyzed using MicroArray Genome Imaging and Clustering Tool (MAGIC Tool) Version 1.0 an open source program. Exposure of E. coli cultures to whey components resulted in transcriptional up regulation of all heat shock related E. coli genes when compared to samples maintained in PBS. Following log transformation three genes were markedly up-regulated as indicated by values greater than 3 for the expression ratio. The gene encoding 2-oxoglutarate dehydrogenase was down regulated (- 2). The differential expression of these genes may serve to identify pathways for the control of E. coli mastitis and indicate targets for intervention.
- Subjects
HIGH Point (N.C.); BOVINE mastitis; GENE expression; WHEY; ESCHERICHIA coli; ESCHERICHIA coli diseases; COMPOSITION of milk
- Publication
Journal of Animal Science, 2006, Vol 84, p309
- ISSN
0021-8812
- Publication type
Article