title: Molecular cloning and characterization of the NADPH oxidase from the kuruma shrimp, Marsupenaeus japonicus: early gene up-regulation after Vibrio penaeicida and poly(I:C) stimulations in vitro.
abstract: Free radicals such as nitric oxide (NO) and reactive oxygen species (ROS) are involved in many physiological processes. In humans, there are 5 homologs of nicotinamide adenine dinucleotide phosphate (NADPH) oxidases (Noxes) that generate superoxide (O(2)(-)), which can dismute to produce ROS, and play significant roles in innate immunity and cell proliferation. Though Noxes have been identified in vertebrates (humans and fishes) and some insects, there are very few reports investigating Noxes in crustaceans. In the present study, we describe the entire cDNA sequence (4216 bp) of Marsupenaeus japonicus (kuruma shrimp) Nox (MjNox) generated using reverse transcriptase-polymerase chain reaction (RT-PCR) and random amplification of cDNA ends (RACE). The open reading frame of MjNox encodes a protein of 1280 amino acids with an estimated mass of 146 kDa that has 46.8% sequence homology with the Nox gene of the fruit fly, Drosophila melanogaster. Highly conserved amino acid sequences were observed in the NADPH binding domain. Transcriptional analysis revealed that MjNox mRNA is highly expressed in the lymphoid organ, hepatopancreas and hemocytes of the healthy kuruma shrimp. In the hemocytes, MjNox expression reached its peak 4 h after stimulation with either Vibrio penaeicida or poly(I:C) and decreased to its normal level after 12 h.This study is the first to identify and clone a Nox family member (MjNox) from a crustacean species.
authors: Interdisciplinary Graduate School of Agriculture and Engineering, University of Miyazaki, 1-1 Gakuen Kibanadai-nishi, 889-2192 Miyazaki, Japan.
authors: Mari
authors: M
authors: Inada
authors: Raja
authors: R
authors: Sudhakaran
authors: Keisuke
authors: K
authors: Kihara
authors: Junichi
authors: J
authors: Nishi
authors: Maki
authors: M
authors: Yoshimine
authors: Tohru
authors: T
authors: Mekata
authors: Tomoya
authors: T
authors: Kono
authors: Masahiro
authors: M
authors: Sakai
authors: Terutoyo
authors: T
authors: Yoshida
authors: Toshiaki
authors: T
authors: Itami
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular / Q000379:methods*; D015971:Gene Expression Regulation, Enzymologic; D006434:Hemocytes / Q000201:enzymology; D043143:Hepatopancreas / Q000201:enzymology; D008221:Lymphoid Tissue / Q000201:enzymology; D008969:Molecular Sequence Data; D019255:NADPH Oxidases / Q000235:genetics* / Q000378:metabolism*; D016366:Open Reading Frames / Q000235:genetics*; D033561:Penaeidae / Q000201:enzymology* / Q000235:genetics / Q000382:microbiology; D010802:Phylogeny; D012333:RNA, Messenger / Q000235:genetics; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D015854:Up-Regulation; D014733:Vibrio / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D019255:NADPH Oxidases