重组T7噬菌体生物学特性的研究  

Study on Biological Characteristics of Recombinant T7 Phage

作　　者：王义伟[1] 徐海[1] 陈瑾[1] 郑其升[1] 侯继波[1] 

机构地区：[1]江苏省农业科学院国家兽用生物制品工程技术研究中心,江苏南京210014

出　　处：《动物医学进展》2014年第1期36-40,共5页Progress In Veterinary Medicine

基　　金：江苏省农业自主创新基金项目[CX(10)216]

摘　　要：以大肠埃希菌BL21为宿主菌研究重组T7噬菌体生物学特性。将构建好的重组T7噬菌体扩增纯化，分析其对PH、温度及离子浓度的敏感性，并将重组噬菌体与宿主菌以不同比例混合测定最佳感染复数并绘制其一步生长曲线。结果表明，重组T7噬菌体的热稳定性较弱，在60℃时20min效价明显下降，在70℃时5min内即会全部失活；PH4～11的范围内重组T7噬菌体均可稳定存在；培养基中Mg件浓度为10mmol／L及Ca2＋浓度为5mmol／L时最有利于噬斑的形成，出斑率提高了约20％；重组噬菌体感染其宿主大肠埃希菌的最佳感染复数为10-5；其一步生长曲线的潜伏期约30min，裂解期约150min，裂解量为126pfu／cell。构建的重组T7噬菌体具有较强的裂解能力，潜伏期短，裂解量大，对理化因素稳定性好，为进一步大规模、高效率制造口蹄疫表位疫苗提供了理论基础。This paper investigated the biological properties of recombinant T7 phages based on E. coli BL21 as host strain. The physiological characteristics such as the optimal multiplicity of infection（MOI）, the one-step growth curves and the sensitivity to pH, temperature and ionic concentration were tested after re- combinant T7 phages were amplified and purified. The results showed that the thermo-stability of recom- binant T7 phages was weak,the titer decreased obviously after heating at 60 ℃ for 20 minutes and heating for 5 minutes at the temperature of 70 ℃ could inactivate it completely. The phage had relatively high sta- bility between pH 4 to 11. The plaque formation of recombinant T7 phages was the best on the medium with addition of 10 mmol/L Mg2＋ and 5 mmol/L Ca2＋ , the numbers of plaques were about 20 higher than that in the medium without Mgz＋ or Ca2＋. The optimal MOI was 10-5. The one-step growth curve of the phage showed that the latent period and the rise period of the phage were about 30 min and 150 min,re- spectively,and the average burst size was about 126 pfu/cell. Recombinant T7 phage has shorter incubation period, apparent burst size, good stability to physical and chemical factors. This study can provide a theoret- ical basis for further large-scale, high-efficiency manufacturing foot-and-mouth disease epitope vaccine.

关 键 词：重组T7噬菌体 疫苗 生物学特性 

分 类 号：R383.24[医药卫生—医学寄生虫学]