Construction of a microarray specific to the chicken immune system: profiling gene expression in B cells after lipopolysaccharide stimulation |
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| Authors: | Aimie J Sarson Leah R Read Hamid R Haghighi Melissa D Lambourne Jennifer T Brisbin Huaijun Zhou and Shayan Sharif |
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| Affiliation: | Department of Pathobiology, University of Guelph, Ontario. |
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| Abstract: | The objective of this study was to profile gene expression in cells of the chicken immune system. A low-density immune-specific microarray was constructed that contained genes with known functions in the chicken immune system, in addition to chicken-expressed sequence tags (ESTs) homologous with mammalian immune system genes, which were systematically characterized by bioinformatic analyses. Genes and ESTs that met the annotation criteria were amplified and placed on a microarray. The microarray contained 84 immune system gene elements. As a means of calibration, the microarray was then used to examine gene expression in chicken B cells after lipopolysaccharide stimulation. Differential gene expression was observed at 6, 12, and 24 h but not at 48 h after stimulation. The results were validated by semiquantitative polymerase chain reaction. The microarray showed a high degree of reproducibility, as demonstrated by intra- and interassay correlation coefficients of 0.97 and 0.95, respectively. Thus, the low-density microarray developed in this study may be used as a tool for monitoring gene expression in the chicken immune system. |
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