Development and validation of an oligonucleotide microarray for immuno-inflammatory genes of ruminants |
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| Authors: | Craig Watkins Annie McKellar Kirsty Jensen Abraham George Doug Jones Michael J Sharp Karen Stevenson John Hopkins |
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| Institution: | (1) Moredun Research Institute, Pentland Science Park, Bush Loan, Penicuik, Midlothian, EH26 0PZ, UK;(2) Roslin Institute &; Royal (Dick) School of Veterinary Studies, University of Edinburgh, Roslin Biocentre, Midlothian, EH25 9PS, UK;(3) Centre for Infectious Disease, Royal (Dick) School of Veterinary Studies (R(D)SVS), University of Edinburgh, Summerhall, Edinburgh, EH9 1QH, UK;(4) Present address: Veterinary Laboratory Agency, Pentland Science Park, Bush Loan, Midlothian, EH26 0PZ, UK; |
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| Abstract: | This report describes the development of small DNA microarrays of fully defined genes suitable for projects requiring detailed
analysis of gene expression in sheep and/or cattle. Two arrays have been developed; the first is a small reference microarray
(RIGRA) that has been used to validate experimental design and methodology; the second, a larger array (RIGUA) containing
probes for 516 ruminant immuno-inflammatory genes, each represented by non-overlapping 75mer oligonucleotides. Experiments
used to validate this microarray were: (1) a comparison of gene expression profiles from sheep broncho-alveolar macrophages
before and after in vitro activation with lipopolysaccharide (LPS), using the RIGRA; (2) the differential gene expression between five in vitro unstimulated sheep keratinocyte cultures; (3) LPS/interferon γ stimulated and unstimulated blood monocytes purified from
Holstein-Friesians (Bos taurus) and Sahiwals (Bos indicus) cattle using the RIGUA. Real-time, quantitative RT-PCR was used to validate the gene expression profiles obtained with the
RIGUA microarrays. The potential for using such an immunological tool in understanding the relative gene expression corresponding
to immune-inflammatory responses of sheep and cattle is discussed.
Supplementary information for this article may be found at (login: watkins and password: PuBm7t) |
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| Keywords: | Ruminant Microarray Immuno-inflammatory Gene expression |
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