Application of disease-associated differentially expressed genes--mining for functional candidate genes for mastitis resistance in cattle.
In this study the mRNA differential display method was applied to identify mastitis-associated expressed DNA sequences based on different expression patterns in mammary gland samples of non-infected and infected udder quarters of a cow. In total, 704 different cDNA bands were displayed in both udder samples. Five hundred-and-thirty two bands, (75.6%) were differentially displayed. Ninety prominent cDNA bands were isolated, re-amplified, cloned and sequenced resulting in 87 different sequences. Amongst the 19 expressed sequence tags showing a similarity with previously described genes, the majority of these sequences exhibited homology to protein kinase encoding genes (26.3%), to genes involved in the regulation of gene expression (26.3%), to growth and differentiation factor encoding genes (21.0%) and to immune response or inflammation marker encoding genes (21.0%). These sequences were shown to have mastitis-associated expression in the udder samples of animals with and without clinical mastitis by quantitative RT-PCR. They were mapped physically using a bovine-hamster somatic cell hybrid panel and a 5000 rad bovine whole genome radiation hybrid panel. According to their localization in QTL regions based on an established integrated marker/gene-map and their disease-associated expression, four genes (AHCY, PRKDC, HNRPU, OSTF1) were suggested as potentially involved in mastitis defense.
Schwerin
Manfred
M
Research Unit for Molecular Biology, Research Institute for the Biology of Farm Animals, Dummerstorf, Germany. schwerin@tbn-dummerstorf.de
Czernek-Schäfer
Diana
D
Goldammer
Tom
T
Kata
Srinivas R
SR
Womack
James E
JE
Pareek
Ravi
R
Pareek
Chandra
C
Walawski
Krzysztof
K
Brunner
Ronald M
RM
D000818:Animals; D002417:Cattle; D002874:Chromosome Mapping; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D020224:Expressed Sequence Tags; D005260:Female; D020869:Gene Expression Profiling; D008414:Mastitis, Bovine / Q000235:genetics* / Q000517:prevention & control; D017385:Sequence Homology
D017931:DNA Primers; D018076:DNA, Complementary