title: Cloning and expression analysis of a novel high-mobility group box 2 homologue from Lampetra japonica.
abstract: High-mobility group box 2 (HMGB2) is a nonhistone architectural protein that plays important roles in many biological processes. In this study, we cloned a homologue of the HMGB2 from the lymphocyte-like cells of Lampetra japonica (L. japonica). Sequence analysis reveals that L. japonica HMGB2 contains two highly conserved motifs and shares more than 70 % identity with the homologues from other vertebrate species. Subsequently, Lj-HMGB2 was subcloned into the pET-28a(+) and pIRES2 AcGFP1-Nuc vector and expressed in Rosetta blue (DE3) and Hela cell lines, respectively. The recombinant L. japonica HMGB2 (rLj-HMGB2) with apparent molecular mass of 22 kDa was further purified by His-Bind affinity chromatography. Real-time quantitative PCR indicates that the expression level of Lj-HMGB2 was particularly up-regulated in intestines after challenged with lipopolysaccharide, while up-regulated in lymphocyte-like cells and heart after challenged with concanavalin A in vivo. In addition, rLj-HMGB2 could induce the generation of proinflammatory mediators in the activated human acute monocytic leukemia cell line (THP1), which suggested that Lj-HMGB2 may participate in the immune response of the lampreys.
authors.affiliation: Institute of Marine Genomics and Proteomics, Liaoning Normal University, Dalian, China.
authors.forename: Pang
authors.initials: P
authors.lastname: Yue
authors.forename: Xiao
authors.initials: X
authors.lastname: Rong
authors.forename: Xue
authors.initials: X
authors.lastname: Zhuang
authors.forename: Huang Jin
authors.initials: HJ
authors.lastname: Sha
authors.forename: Jin Min
authors.initials: JM
authors.lastname: Li
authors.forename: Liu
authors.initials: L
authors.lastname: Xin
authors.forename: Qing Wei
authors.initials: QW
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004247:DNA / Q000235:genetics; D029941:Fish Proteins / Q000235:genetics* / Q000276:immunology; D015870:Gene Expression; D024261:HMGB2 Protein / Q000235:genetics* / Q000276:immunology; D006367:HeLa Cells; D006801:Humans; D018836:Inflammation Mediators / Q000378:metabolism; D007798:Lampreys / Q000235:genetics* / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology; D008969:Molecular Sequence Data; D009000:Monocytes / Q000276:immunology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis
chemical_list: D029941:Fish Proteins; D024261:HMGB2 Protein; D018836:Inflammation Mediators; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D004247:DNA