吉林农业大学学报Issue(2)：211-215,220,6.DOI:10.13327/j.jjlau.2015.2522
一种大肠杆菌噬菌体新裂解酶的表达与活性检测∗
Expression and Activity Determination of a Novel E. coli Phage Lyase
摘要
Abstract
An E. coli phage vB_EcoM⁃ep3 with good lysis ability was isolated in early experiments. We found an unreported lyase gene(Lysep3)could encode a novel lyase in E. coli phage showing high homology with P. aeruginosa phage lyase after the sequence analysis. To explore lysis activity of Lysep3 in P. aeruginosa and E. coli, gene cloning, prokaryotic expression, protein purification and lysis activity determination in vitro were performed. Results showed that length of lyase gene is 492 bp and protein molecular mass is 17�7 ku. When using Lysep3 (0�5 mg/mL) alone, P. aerugi⁃nosa and E. coli were not lysed while growth could be significantly inhibited. When using Lysep3 and citric acid (0�5 mmol/L) in combination, better lysis ability was displayed ( with continual effect from 2 h to 24 h) ,and quantity of bacteria decreased 7�2 times and 17�3 times respectively. Results suggested that the evolutionary relationship of lyase between vB_EcoM⁃ep3 and P. aeruginosaare re⁃presented in both gene sequence and function. These results not only provide strong evidence for the study on relationship between structure and function of lysine, but also contribute to the transforma⁃tion of a broad spectrum lyase.关键词
噬菌体裂解酶/大肠杆菌/绿脓杆菌/原核表达Key words
phage lyase/Escherichia coli(E. coli)/Pseudomonas aeruginosa(P. aeruginosa)/pro-karyotic expression分类
农业科技引用本文复制引用
吕萌,王爽,闫广谋,冯新,顾敬敏,雷连成..一种大肠杆菌噬菌体新裂解酶的表达与活性检测∗[J].吉林农业大学学报,2015,(2):211-215,220,6.基金项目
国家重点基础研究发展计划项目 （）
