title: Molecular cloning, characterization and expression analysis of trypsin-like serine protease from triangle-shell pearl mussel (Hyriopsis cumingii).
abstract: Trypsin-like serine protease (TLS) is ubiquitous in animals and plays a number of diverse roles, including dietary protein digestion, hemolymph coagulation, antimicrobial activity and immune responses, among others. This study reports the isolation of a 1048 bp full-length cDNA sequence of TLS from triangle-shell pearl mussel (Hyriopsis cumingii), including a 12 bp 5' UTR (untranslated region), a 172 bp 3' UTR, and an open reading frame (ORF) of 864 bp by rapid amplification of cDNA ends (RACE). Bioinformatic analysis shows that the gene belongs to the trypsin-like serine protease superfamily, and contains a 15 residues N-terminal signal peptide and a conserved C-terminal domain. In comparison to other serine proteases, the catalytic triad were identified as His-98, Asp-149, and Ser-240. Quantitative real-time PCR (qPCR) showed a broad expression of the TLS gene in ten tested tissues. Time-course expression analysis demonstrated that the expression level of the TLS mRNA was significantly up-regulated in eight tested tissues (liver, intestine, gill, heart, axe foot, adductor muscle, kidney and gonad), but down-regulated in mantle and stomach after Aeromonas hydrophila injection. This is one of the results indicate that TLS may be involved in innate defense reactions against A. hydrophila in triangle-shell pearl mussel.
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Hongquan
authors.initials: H
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Liang
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Yurong
authors.initials: Y
authors.lastname: Zhao
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Qiaolin
authors.initials: Q
authors.lastname: Liu
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Yaoguo
authors.initials: Y
authors.lastname: Li
authors.affiliation: College of Bioscience and Biotechnology, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Zili
authors.initials: Z
authors.lastname: Yi
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, 410128, China.
authors.forename: Kaijian
authors.initials: K
authors.lastname: Chen
authors.affiliation: College of Animal Science and Technology, Hunan Agricultural University, Changsha, 410128, China. Electronic address: tyx1128@aliyun.com.
authors.forename: Tiaoyi
authors.initials: T
authors.lastname: Xiao
mesh_terms: D016980:Aeromonas hydrophila / Q000502:physiology; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D007113:Immunity, Innate; D008969:Molecular Sequence Data; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D014018:Tissue Distribution; D049876:Unionidae / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism / Q000382:microbiology
chemical_list: D018076:DNA, Complementary; D012333:RNA, Messenger; C114880:trypsin-like serine protease; D012697:Serine Endopeptidases
keywords: Quantitative real-time PCR; Time-course expression; Triangle-shell pearl mussel; Trypsin-like serine protease