Anti-disease prepn. contg. piginterleukin-4,6 fused gene
A technology of interleukin and fusion gene, which is applied in the field of porcine interleukin-4 and 6 fusion gene anti-disease preparations, can solve the problems that restrict the prevention and treatment of infectious diseases
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[0095] 1. Cloning and sequence analysis of porcine interleukin-4 gene
[0096] Sterile blood was collected from the anterior vena cava of pigs, and the lymphocyte separation technology was used to separate the peripheral blood lymphocytes of pigs for in vitro culture. After culturing for 36 hours under the stimulation of Con A, the total RNA of the cultured lymphocytes was extracted, and the total RNA of Tibetan pigs was extracted. A pair of primers for porcine IL-4 gene cDNA amplification was designed. The primer sequences are as follows:
[0097]Primer 1: CGG GAT CCC TAT TCA TGG GTC TCA CCT C
[0098] Primer 2: CGG GAT CCT CAG CTT CAA CAC TTT
[0099] Using total RNA as a template, add 5 μl of 10×RT-PCR reaction buffer, 25mM MgCl to the 50 μl reaction system 2 10μl, 10mM dNTP5μl, RNase Inhibitor (40units / μl) 1μl, AMV RNase XL (5units / uL) 1μl, AMV-Optimized Taq (5units / uL) 1μl, 5′ and 3′ primers 20uM each, total RNA 1~2μl, water to 50 μl. The RT-PCR cycle parameters are:...
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