Molecular cloning and expression analysis of porcine gamma-interferon-inducible lysosomal thiol reductase (GILT)

A porcine interferon-gamma-inducible lysosomal thiol reductase (GILT) cDNA, designated pGILT, was cloned by RT-PCR and rapid amplification of cDNA ends (RACE) strategies. The full-length cDNA of pGILT consists of 1,062 bp with a 741 bp open reading frame, encoding 246 amino acids, with a putative molecular weight of 29.5 kDa. The deduced pGILT possesses the typical structural feature of mammalian GILT, including an active-site CXXC motif, a GILT signature sequence CQHGX(2)ECX(2)NX(4)C, and 10 conserved cysteines. The genomic DNA sequence of pGILT contains seven exons and six introns, which is similar to vertebrate GILT exon-intron organization. The result of real-time PCR showed that GILT is expressed in many tissues in the pig, including spleen, liver, lung, heart, intestine, blood and kidney. And the pGILT expression is obviously up-regulated in spleen and blood after induction with LPS. These results suggesting that pGILT is highly likely to play a role in the innate immune responses in porcine. It also provided the basis for investigations on the role of GILT in this important domestic species and an animal model for human diseases.