Molecular Cloning and Bioinformatics Analysis of kdpD Gene from Vibrio alginolyticus

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In this study,histidine kinase gene kdpD in the two-component regulatory system of Vibrio alginolyticus strain HY9901 was cloned for bioinformatics analysis. Sequence analysis revealed that kdpD gene( GenB ank accession number: KJ544668) was 1 374 bp in length,encoding a putative protein of 457 amino acids. The predicted molecular weight( MW) of KdpE was 51. 60 kD with a theoretical isoelectric point( pI) of 6. 02. Using SignalP 4. 0,TMHMM Server 2. 0 and SoftB erry-Psite software,it was predicted that KdpE protein was equally located in Golgi apparatus,plasma membrane and endoplasmic reticulum( 33. 3%),which did not contain a signal peptide but contained three transmembrane domains. KdpE protein had five casein kinase II phosphorylation sites,five protein kinase C phosphorylation sites and one cA MP- and cG MP-dependent protein kinase phosphorylation site. A phylogenetic tree was constructed by MEGA 5. 0 software,which revealed that KdpD from V. alginolyticus had close genetic relationship with corresponding proteins from V. campbellii and V. parahaemolyticus. Using SWISSMODEL Workspace,the three-dimensional structure of HATPase_c conserved domain in KdpD protein was constructed. These results may provide the basis for further studies on two-component regulatory system KdpD/KdpE of V. alginolyticus. In this study, histidine kinase gene kdpD in the two-component regulatory system of Vibrio alginolyticus strain HY9901 was cloned for bioinformatics analysis. Sequence analysis revealed that kdpD gene (GenBank accession number: KJ544668) was 1 374 bp in length, encoding a putative The predicted molecular weight (MW) of KdpE was 51. 60 kD with a theoretical isoelectric point (pI) of 6. 02. Using SignalP 4. 0, TMHMM Server 2. 0 and SoftBerry-Psite software , it was predicted that KdpE protein was equally located in Golgi apparatus, plasma membrane and endoplasmic reticulum (33.3%), which did not contain a signal peptide but contained three transmembrane domains. KdpE protein had five casein kinase II phosphorylation sites, five protein kinase C phosphorylation sites and one cA MP- and cG MP-dependent protein kinase phosphorylation site. A phylogenetic tree was constructed by MEGA 5. 0 software, which revealed that KdpD from V. alginolyticus had close genetic relatio nship with corresponding proteins from V. campbellii and V. parahaemolyticus. Using SWISSMODEL Workspace, the three-dimensional structure of HATPase_c conserved domain in KdpD protein was constructed. These results may provide the basis for further studies on two-component regulatory system KdpD / KdpE of V. alginolyticus.
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