title: Gene profiling and characterization of arginine kinase-1 (MrAK-1) from freshwater giant prawn (Macrobrachium rosenbergii).
abstract: Arginine kinase-1 (MrAK-1) was sequenced from the freshwater prawn Macrobrachium rosenbergii using Illumina Solexa Genome Analyzer Technique. MrAK-1 consisted of 1068 bp nucleotide encoded 355 polypeptide with an estimated molecular mass of 40 kDa. MrAK-1 sequence contains a potential ATP:guanido phosphotransferases active domain site. The deduced amino acid sequence of MrAK-1 was compared with other 7 homologous arginine kinase (AK) and showed the highest identity (96%) with AK-1 from cherry shrimp Neocaridina denticulate. The qRT-PCR analysis revealed a broad expression of MrAK-1 with the highest expression in the muscle and the lowest in the eyestalk. The expression of MrAK-1 after challenge with the infectious hypodermal and hematopoietic necrosis virus (IHHNV) was tested in muscle. In addition, MrAK-1 was expressed in Escherichia coli by prokaryotic expression plasmid pMAL-c2x. The optimum temperature (30 °C) and pH (8.5) was determined for the enzyme activity assay. MrAK-1 showed significant (P < 0.05) activity towards 10-50 mM ATP concentration. The enzyme activity was inhibited by α-ketoglutarate, glucose and ATP at the concentration of 10, 50 and 100 mM respectively. Conclusively, the findings of this study indicated that MrAK-1 might play an important role in the coupling of energy production and utilization and the immune response in shrimps.
authors.affiliation: Centre for Biotechnology in Agriculture Research, Division of Genetics & Molecular Biology, Institute of Biological Sciences, Faculty of Science, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors.forename: Jesu
authors.initials: J
authors.lastname: Arockiaraj
authors.forename: Puganeshwaran
authors.initials: P
authors.lastname: Vanaraja
authors.forename: Sarasvathi
authors.initials: S
authors.lastname: Easwvaran
authors.forename: Arun
authors.initials: A
authors.lastname: Singh
authors.forename: Tahereh
authors.initials: T
authors.lastname: Alinejaid
authors.forename: Rofina Yasmin
authors.initials: RY
authors.lastname: Othman
authors.forename: Subha
authors.initials: S
authors.lastname: Bhassu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001122:Arginine Kinase / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D019223:Densovirinae / Q000502:physiology*; D004734:Energy Metabolism; D004926:Escherichia coli / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D008969:Molecular Sequence Data; D009132:Muscles / Q000378:metabolism; D033541:Palaemonidae / Q000145:classification / Q000235:genetics* / Q000276:immunology / Q000821:virology; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
chemical_list: D001122:Arginine Kinase