title: Cloning, expression, characterization and phylogenetic analysis of arginine kinase from greasyback shrimp (Metapenaeus ensis).
abstract: Arginine kinase (AK) plays an important role in cellular energy metabolism in invertebrate. The encoding AK gene from Shrimp Metapenaeus ensis (M. ensis) was cloned in prokaryotic expression plasmid pET-28a, and it was then expressed in Escherichia coil in dissoluble form. The recombinant protein was purified by following three chromatography steps in turn: CM-Cellulose cation-exchange, Sephacryl S-100HR gel filtrate and DEAE-Sepharose anion-exchange. The purified AK's apparent K(m) was 2.33+/-0.1 and 1.59+/-0.2 mM for ATP and l-arginine, respectively, while its optimum pH and temperature was 8.5 and 30 degrees C in the process of forward reaction, respectively. Phylogenetic analysis of cDNA-derived amino acid sequences for the AKs indicated a close affinity of M. ensis and another shrimp (Litopenaeus vannamei).
authors.affiliation: State Key Laboratory of Virology, College of Life Sciences, Center of Nanoscience and Nanotechnology, Wuhan University, Wuhan, Hubei 430072, PR China.
authors.forename: Jin-Song
authors.initials: JS
authors.lastname: Wang
authors.forename: Zhong-Liang
authors.initials: ZL
authors.lastname: Zheng
authors.forename: Jing
authors.initials: J
authors.lastname: Lei
authors.forename: Ji-Cheng
authors.initials: JC
authors.lastname: Pan
authors.forename: Guo-Lin
authors.initials: GL
authors.lastname: Zou