Expression and functional study of VpV262 Pol,a moderately halophilic DNA polymerase from the Vibrio parahaemolyticus phage VpV262
作者单位:Research Center of Molecular Diagnostics and SequencingResearch Institute of Tsinghua University in Shenzhen MOE Key Lab of BioinformaticsSchool of Life SciencesTsinghua University
会议名称:《中国生物化学与分子生物学会2020全国学术在线会议》
会议日期:2020年
学科分类:1007[医学-药学(可授医学、理学学位)] 100705[医学-微生物与生化药学] 1001[医学-基础医学(可授医学、理学学位)] 100103[医学-病原生物学] 10[医学]
关 键 词:DNA polymerase salt tolerance halophilic enzyme Vibrio parahaemolyticus phage VpV262 3D-Structure random coil
摘 要:Halophilic organisms are found widely in environments where the salt concentration is higher than 0.2 *** proteins isolated from these organisms maintain structural integrity and function under high salt stress,whereas their non-halophilic homologs tend to aggregate and *** we report for the first time the expression and function of a DNA polymerase(DNAPol) VpV262 Pol,which belongs to DNAPol Family A from Vibrio parahaemolyticus phage *** activity assay revealed that VpV262 Pol possessed 5’-3’ polymerase activity as well as 3’-5’ proofreading exonuclease ***262 Pol requires Mg or Mn to catalyze the polymerization *** activity assay under a wide range of salt concentrations showed that VpV262 Pol maintains the highest polymerase activity with 0-0.3 M of NaCl/KCl and0-0.5 M KAc(potassium acetate)/KGlc(potassium gluconate) when treated with 0-1 M corresponding salts,in contrast to significantly decreased activity of Phi29 Pol and Taq Pol above 0.2 *** with typical features of other halophilic proteins,negatively-charged amino acids are more frequently distributed on the surface of VpV262 Pol,contributing to highly solubility and enhanced *** 3 D-Structure of VpV262 Pol needs to be confirmed by experimental data further,this study here has added a member for the relatively small family of halotolerant DNA polymerase,and provides a valuable reference in isolation and characterization of DNA polymerases from halophilic organisms.