Identification of IL-1α gene from bovine peripheral blood mononuclear cells by polymerase chain reaction

Abstract

Interleukin-1 (IL-1) play its role in inflammation, fever and release of acute phase proteins by acting on major organs of the body. IL- 1α gene along with other cytokines compels Th1 and Th17 inflammatory responses. The present study is aimed at standardization of polymerase chain reaction for amplification of IL-1α gene from bovine peripheral blood mononuclear cells. Blood samples were collected in containers with 0.1% Ethylenediaminetetraacetic acid (EDTA) anticoagulant (Sigma, USA). Peripheral blood mononuclear cells (PBMCs) were isolated using commercially available Histopaque®-1077. Extracted total ribose nucleic acid (RNA) was utilized in reverse transcription to prepare complementary deoxyribose nucleic acid (cDNA). IL-1α gene specific primers were used to amlify partial nucleotide sequence of IL-1α. The cDNA was employed in polymerase chain reaction (PCR). The resulting IL-1α DNA product (124 bp) was identified on agar gel electrophoresis and documented.