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Genetics and Molecular Biology
On-line version ISSN 1678-4685
ZAROS, Lilian Giotto; BRICARELLO, Patrízia Ana; AMARANTE, Alessandro Francisco Talamini and COUTINHO, Luiz Lehmann. Quantification of bovine cytokine gene expression using real-time RT-PCR methodology. Genet. Mol. Biol. [online]. 2007, vol.30, n.3, pp. 575-579. ISSN 1678-4685. http://dx.doi.org/10.1590/S1415-47572007000400012.
T cells produce cytokines that affect host response to infection. This paper reports real-time RT-PCR conditions and validation steps for accurate quantification of Bos indicus cytokines, interleukin (IL)-2, IL-4, IL-8, IL12p-35, IL-13, tumoral necrosis factor (TNF)-a, interferon (IFN)-g, monocyte chemoattractant proteins (MCP)-1 and MCP-2, and the glycoprotein mucin (MUC)-1 in two groups of Nelore cattle, one resistant and the other susceptible to gastrointestinal nematode infections. RPL-19 was shown to be an ideal internal control gene, since its expression was constant across treatments and presented lower variation when compared to the GAPDH gene. The optimized conditions established in the present study can be used to determine the immune response of cattle under different experimental conditions, such as viral, bacterial and parasite infections.
Keywords : cytokine gene expression; real-time RT-PCR; cattle; gastrointestinal nematodes; bovine cytokines.