Molecular cloning, expression and characterization of an interleukin 27 p28 homolog in pig (Sus scrofa).
IL-27 is the newest member of the IL-12 cytokine family and plays an important role in the immune regulation. It is composed of two subunits, p28 and EBV-induced gene 3(EBI3). Although human and mouse IL-27 p28 genes have been cloned, pig IL-27 p28 gene has not ever been reported. In the present study, we have cloned and characterized the full-length cDNA of IL-27 p28 from pig. The open reading frame of pig IL-27 p28 gene is 720 bp, which encodes a protein of 239 amino acids with a predicted molecular mass of 26.6 kDa. The deduced amino acid sequence of pig IL-27 p28 showed a high degree of homology to human (63%) and mouse (58%). It was a 4-helix cytokine and belonged to 4-helix cytokine superfamily. Pig IL-27 p28 has one transmembrane region, one signal peptide, and one N-glycosylation site, two Protein kinase C phosphorylation sites, three Casein kinase II phosphorylation sites and one N-myristoylation site. For the expression of pig IL-27 p28 protein in a eukaryotic expression system the recombinant plasmid was constructed. The expression of pig IL-27 p28 in mammalian cells were confirmed by flow cytometry analysis, immunofluorescence and Western blot. The analysis also confirmed a cross reactivity with anti-mouse IL-27 p28 antibody. This is the first report of cloning and characterization of IL-27 p28 in pig.
Hu
Peng
P
Institute for Viral Hepatitis, Key Laboratory of Molecular Biology for Infectious Diseases, Ministry of Education, The Second Affiliated Hospital, Chongqing Medical University, 74# Linjiang Road, Chongqing, China. hp_cq@yahoo.com.cn
Peng
Ming-Li
ML
Chen
Min
M
Hu
Huai-Dong
HD
Tang
Li
L
Tang
Kai-Fu
KF
Ren
Hong
H
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D002478:Cells, Cultured; D003001:Cloning, Molecular; D003429:Cross Reactions; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D007378:Interleukins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D016366:Open Reading Frames; D010802:Phylogeny; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity; D034421:Sus scrofa / Q000235:genetics* / Q000276:immunology*
D017931:DNA Primers; D018076:DNA, Complementary; C486767:Il27 protein, mouse; D007378:Interleukins; C464565:MYDGF protein, human; D021122:Protein Subunits; D011994:Recombinant Proteins