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Construction and validation of a Bovine Innate Immune Microarray.
- Source :
-
BMC genomics [BMC Genomics] 2005 Sep 22; Vol. 6, pp. 135. Date of Electronic Publication: 2005 Sep 22. - Publication Year :
- 2005
-
Abstract
- Background: Microarray transcript profiling has the potential to illuminate the molecular processes that are involved in the responses of cattle to disease challenges. This knowledge may allow the development of strategies that exploit these genes to enhance resistance to disease in an individual or animal population.<br />Results: The Bovine Innate Immune Microarray developed in this study consists of 1480 characterised genes identified by literature searches, 31 positive and negative control elements and 5376 cDNAs derived from subtracted and normalised libraries. The cDNA libraries were produced from 'challenged' bovine epithelial and leukocyte cells. The microarray was found to have a limit of detection of 1 pg/microg of total RNA and a mean slide-to-slide correlation co-efficient of 0.88. The profiles of differentially expressed genes from Concanavalin A (ConA) stimulated bovine peripheral blood lymphocytes were determined. Three distinct profiles highlighted 19 genes that were rapidly up-regulated within 30 minutes and returned to basal levels by 24 h; 76 genes that were up-regulated between 2-8 hours and sustained high levels of expression until 24 h and 10 genes that were down-regulated. Quantitative real-time RT-PCR on selected genes was used to confirm the results from the microarray analysis. The results indicate that there is a dynamic process involving gene activation and regulatory mechanisms re-establishing homeostasis in the ConA activated lymphocytes. The Bovine Innate Immune Microarray was also used to determine the cross-species hybridisation capabilities of an ovine PBL sample.<br />Conclusion: The Bovine Innate Immune Microarray has been developed which contains a set of well-characterised genes and anonymous cDNAs from a number of different bovine cell types. The microarray can be used to determine the gene expression profiles underlying innate immune responses in cattle and sheep.
- Subjects :
- Animals
Cattle
Concanavalin A chemistry
DNA, Complementary metabolism
Down-Regulation
Gene Expression Profiling
Gene Expression Regulation
Gene Library
Immunity
Lymphocytes metabolism
Models, Biological
Models, Statistical
Nucleic Acid Hybridization
Phosphorylation
RNA, Messenger metabolism
Reverse Transcriptase Polymerase Chain Reaction
Sheep
Staphylococcus aureus metabolism
Time Factors
Transcriptional Activation
Up-Regulation
Immune System metabolism
Oligonucleotide Array Sequence Analysis methods
Subjects
Details
- Language :
- English
- ISSN :
- 1471-2164
- Volume :
- 6
- Database :
- MEDLINE
- Journal :
- BMC genomics
- Publication Type :
- Academic Journal
- Accession number :
- 16176586
- Full Text :
- https://doi.org/10.1186/1471-2164-6-135