Open AccessCloning, in vitro expression, and bioactivity of interleukin‐18 isolated from a domestic porcine breed found in Henan
Author(s) -
Chen HongYing,
Zheng LanLan,
Li XinSheng,
Wei ZhanYong,
Cui BaoAn,
Li XiaoKang,
Liu JinPeng,
Yin HongZheng,
Meng JiangTao,
Zhang Yan,
Li ShiMin
Publication year - 2009
Publication title -
fems immunology & medical microbiology
Language(s) - English
Resource type - Journals
eISSN - 1574-695X
pISSN - 0928-8244
DOI - 10.1111/j.1574-695x.2009.00589.x
Subject(s) - biology , microbiology and biotechnology , porcine reproductive and respiratory syndrome virus , porcine parvovirus , population , fusion protein , amino acid , virology , recombinant dna , virus , gene , biochemistry , demography , sociology
To evaluate the effects of recombinant porcine interleukin‐18 (rpIL‐18) on the replication of viruses in host cells and proliferation of lymphocytes, porcine IL‐18 ( pIL‐18 ) isolated from a domestic big‐white porcine breed found in the Henan province (HN) was cloned using a reverse transcriptase‐PCR. The cloned HN pIL‐18 contained an ORF of 579 base pairs encoding a 192‐amino‐acid precursor protein. The amino acid sequence of HN pIL‐18 was compared with all the other pIL‐18 amino acid sequences and varied by at least one amino acid to the consensus of all the others available. HN pIL‐18 mature protein gene was inserted into a prokaryotic vector pGEX‐4T‐1 and expressed in Escherichia coli BL21. The expression of glutathione‐ S ‐transferase–pIL18 fusion protein was confirmed by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis and Western blot analysis. The rpIL‐18 induced in vitro proliferation of concanavalin‐A‐stimulated porcine splenocytes, as revealed by the MTT assay. We studied the antiviral activities of the rpIL‐18 on the replication of porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), and porcine parvovirus (PPV) cultured in two homologous cell lines. The results suggested that rpIL‐18 can stimulate the proliferation of lymphocytes and inhibit viral pathogens infecting the porcine population.