Cloning, genetic analysis and baculovirus expression vector construction of porcine interleukin-2

One pair of primers was designed according to porcine IL-2 gene sequences published in GenBank.The interleukin-2 cDNA of porcine was amplified by RT-PCR from total RNA extracted from spleen lymphocytes stimulated with 10 mg/L ConA in vitro for 27 hours.Then the amplified cDNA was cloned into pGEM-T Easy vector and sequenced.The result showed that the full length of the porcine IL-2 was 516 bp(ORF was 465 bp),encoding a peptide of 154 amino acids whose molecular is 17 400,pI is 5.27.The sequence was just the same as GenBank published.Compared of nhe nucleotides sequence of porcine IL-2 with those of cat,cattle,chicken,dog,duck,goat,horse,human and house mouse.The result showed that the nucleotides homologies were 83.7%,82.6%,28.2%,80.6%,29.6%,83.4%,81.3%,82.0% and 61.5%,respectively.The baculovirus transfer vector of pFastBacDual containing pIL-2 was constructed and transformed into DH10Bac which contains a shuttle vector of bacmid.The study paved the way of the expression in insect cell and the development of new immunologic adjuvant.