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.