[Objectives]To amplify the DNA-binding response regulator PhoP in Vibrio alginolyticus and analyze its sequence characteristics and subunit structure.[Methods]According to the sequence of the DNA-binding response regu...[Objectives]To amplify the DNA-binding response regulator PhoP in Vibrio alginolyticus and analyze its sequence characteristics and subunit structure.[Methods]According to the sequence of the DNA-binding response regulator PhoP in V.alginolyticus,a pair of specific primers was designed for PCR amplification,and the bioinformatics of the sequence amplified was analyzed.Using MEGA 5.0 software,the phoP phylogenetic tree was constructed by the neighbor-joining method.Using SWISS-MODEL software,the three-dimensional structural model of the PhoP subunit was simulated.[Results]The full-length phoP gene was 732 bp,encoding a total of 243 amino acids.The predicted theoretical molecular weight of the protein is about 27.67 kD,and the isoelectric point is 5.09.The prediction results of protein subcellular localization,SignalP 4.0,TMHMM Server 2.0 and SoftBerry-Psite show that PhoP is located in the cytoplasm,and is stable and hydrophobic;there is a signal peptide cleavage site between amino acids 29 and 30,and there is no transmembrane region.The amino acid sequence contains one Asn-glycosylation site,one protein kinase C phosphorylation site,seven casein kinase II phosphorylation sites,one tyrosine kinase phosphorylation site,three myristoylation sites,and seven C-terminal microbody targeting signal sites.The PhoP of V.alginolyticus has high homology with that of Vibrio campbellii.The PhoP subunit of V.alginolyticus has similar configuration to the single-subunit RegX3 protein of Mycobacterium tuberculosis.[Conclusions]This study has a positive effect on the prevention and control of vibriosis and the improvement of the current aquatic economic animal breeding environment.展开更多
基金Outstanding Graduate Entering Laboratory Project of College of Fisheries,Guangdong Ocean UniversityNational Natural Science Foundation of China(32073015)+3 种基金Natural Science Foundation of Guangdong Province(2021A1515011078)Special Fund for Science and Technology Innovation Strategy of Guangdong Province(Undergraduate Science and Technology Innovation Cultivation)(pdjh2021b0239)Undergraduate Innovation and Entrepreneurship Training Program of Guangdong Ocean University(CXXL2021122)Undergraduate Innovation Team of Guangdong Ocean University(CCTD-201802).
文摘[Objectives]To amplify the DNA-binding response regulator PhoP in Vibrio alginolyticus and analyze its sequence characteristics and subunit structure.[Methods]According to the sequence of the DNA-binding response regulator PhoP in V.alginolyticus,a pair of specific primers was designed for PCR amplification,and the bioinformatics of the sequence amplified was analyzed.Using MEGA 5.0 software,the phoP phylogenetic tree was constructed by the neighbor-joining method.Using SWISS-MODEL software,the three-dimensional structural model of the PhoP subunit was simulated.[Results]The full-length phoP gene was 732 bp,encoding a total of 243 amino acids.The predicted theoretical molecular weight of the protein is about 27.67 kD,and the isoelectric point is 5.09.The prediction results of protein subcellular localization,SignalP 4.0,TMHMM Server 2.0 and SoftBerry-Psite show that PhoP is located in the cytoplasm,and is stable and hydrophobic;there is a signal peptide cleavage site between amino acids 29 and 30,and there is no transmembrane region.The amino acid sequence contains one Asn-glycosylation site,one protein kinase C phosphorylation site,seven casein kinase II phosphorylation sites,one tyrosine kinase phosphorylation site,three myristoylation sites,and seven C-terminal microbody targeting signal sites.The PhoP of V.alginolyticus has high homology with that of Vibrio campbellii.The PhoP subunit of V.alginolyticus has similar configuration to the single-subunit RegX3 protein of Mycobacterium tuberculosis.[Conclusions]This study has a positive effect on the prevention and control of vibriosis and the improvement of the current aquatic economic animal breeding environment.
