Construction of a microarray specific to the chicken immune system: profiling gene expression in B cells after lipopolysaccharide stimulation.
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.
Sarson
Aimie J
AJ
Department of Pathobiology, University of Guelph, Ontario.
Read
Leah R
LR
Haghighi
Hamid R
HR
Lambourne
Melissa D
MD
Brisbin
Jennifer T
JT
Zhou
Huaijun
H
Sharif
Shayan
S
D000818:Animals; D001402:B-Lymphocytes / Q000276:immunology*; D001483:Base Sequence; D002645:Chickens / Q000235:genetics* / Q000276:immunology*; D020869:Gene Expression Profiling / Q000379:methods / Q000662:veterinary; D005786:Gene Expression Regulation; D007107:Immune System / Q000166:cytology / Q000276:immunology; D008070:Lipopolysaccharides / Q000494:pharmacology*; D046228:Microarray Analysis / Q000379:methods / Q000662:veterinary*; D008969:Molecular Sequence Data; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary
D008070:Lipopolysaccharides