title: Cloning and characterization of two duplicated interleukin-17A/F2 genes in common carp (Cyprinus carpio L.): Transcripts expression and bioactivity of recombinant IL-17A/F2.
abstract: Interleukin-17 (IL-17) plays an important role in inflammation and host defense in mammals. In this study, we identified two duplicated IL-17A/F2 genes in the common carp (Cyprinus carpio) (ccIL-17A/F2a and ccIL-17A/F2b), putative encoded proteins contain 140 amino acids (aa) with conserved IL-17 family motifs. Expression analysis revealed high constitutive expression of ccIL-17A/F2s in mucosal tissues, including gill, skin and intestine, their expression could be induced by Aeromonas hydrophila, suggesting a potential role in mucosal immunity. Recombinant ccIL-17A/F2a protein (rccIL-17A/F2a) produced in Escherichia coli could induce the expression of proinflammatory cytokines (IL-1β) and the antimicrobial peptides S100A1, S100A10a and S100A10b in the primary kidney in a dose- and time-dependent manner. Above findings suggest that ccIL-17A/F2 plays an important role in both proinflammatory and innate immunity. Two duplicated ccIL-17A/F2s showed different expression level with ccIL-17A/F2a higher than b, comparison of two 5' regulatory regions indicated the length from anticipated promoter to transcriptional start site (TSS) and putative transcription factor binding site (TFBS) were different. Promoter activity of ccIL-17A/F2a was 2.5 times of ccIL-17A/F2b which consistent with expression results of two genes. These suggest mutations in 5'regulatory region contributed to the differentiation of duplicated genes. To our knowledge, this is the first report to analyze 5'regulatory region of piscine IL-17 family genes.
authors.affiliation: Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China. Electronic address: lihx@ffrc.cn.
authors.forename: Hongxia
authors.initials: H
authors.lastname: Li
authors.affiliation: Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China; Wuxi Fishery College, Nanjing Agricultural University, Wuxi 214081, China. Electronic address: yujh@ffrc.cn.
authors.forename: Juhua
authors.initials: J
authors.lastname: Yu
authors.affiliation: Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China. Electronic address: lijl@ffrc.cn.
authors.forename: Jianlin
authors.initials: J
authors.lastname: Li
authors.affiliation: Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China. Electronic address: tangyk@ffrc.cn.
authors.forename: Yongkai
authors.initials: Y
authors.lastname: Tang
authors.affiliation: Key Laboratory of Freshwater Fisheries and Germplasm Resources Utilization, Ministry of Agriculture, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China. Electronic address: yuf@ffrc.cn.
authors.forename: Fan
authors.initials: F
authors.lastname: Yu
authors.affiliation: Wuxi Fishery College, Nanjing Agricultural University, Wuxi 214081, China. Electronic address: 714203849@qq.com.
authors.forename: Jie
authors.initials: J
authors.lastname: Zhou
authors.affiliation: Wuxi Fishery College, Nanjing Agricultural University, Wuxi 214081, China. Electronic address: 249545226@qq.com.
authors.forename: Wenjuan
authors.initials: W
authors.lastname: Yu
mesh_terms: D000249:Adenosine Monophosphate / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D002347:Carps / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D005880:Gills / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D020381:Interleukin-17 / Q000235:genetics* / Q000378:metabolism; D053583:Interleukin-1beta / Q000378:metabolism; D007413:Intestinal Mucosa / Q000378:metabolism; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D010802:Phylogeny; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D012867:Skin / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D018076:DNA, Complementary; D020381:Interleukin-17; D053583:Interleukin-1beta; D012333:RNA, Messenger; D011994:Recombinant Proteins; D000249:Adenosine Monophosphate
keywords: Common carp (Cyprinus carpio); Duplicated genes; Expression; IL-17A/F2; Proinflammatory and innate immune function; Promoter activity