[1]张远星,李统战,余子豪,等.三种噬菌体裂解基因对大肠杆菌致死效果的比较[J].中国预防兽医学报,2019,(12):1268-1274.[doi:0.3969/j.issn.1008-0589.201903025]
 ZHANG Yuan-xing,LI Tong-zhan,YU Zi-hao,et al.Lethality of three phage lysis genes to host Escherichia coli strains[J].Chinese journal of preventive veterinary medicine,2019,(12):1268-1274.[doi:0.3969/j.issn.1008-0589.201903025]
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三种噬菌体裂解基因对大肠杆菌致死效果的比较()
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《中国预防兽医学报》[ISSN:1008-0589/CN:23-1417/S]

卷:
期数:
2019年12
页码:
1268-1274
栏目:
免疫学
出版日期:
2020-01-25

文章信息/Info

Title:
Lethality of three phage lysis genes to host Escherichia coli strains
文章编号:
1008-0589(2019)12-1268-07
作者:
 

张远星李统战余子豪查 帆李倩文余旭平*

 (浙江大学 动物科学学院,浙江 杭州 310058)
Author(s):
 

ZHANG Yuan-xing LI Tong-zhan YU Zi-hao ZHA Fan LI Qian-wen YU Xu-ping*

 (College of Animal Science, Zhejiang University, Hangzhou 310058, China)
关键词:
噬菌体裂解基因过表达致死性
Keywords:
 bacteriophage lysis gene  over-expression  lethality
分类号:
S852.61
DOI:
0.3969/j.issn.1008-0589.201903025
文献标志码:
A
摘要:
为比较3种噬菌体裂解基因M-lysis、mE和E对大肠杆菌的致死效果,本研究将噬菌体裂解基因M-lysis、mE和E克隆于严谨调控的高效表达载体pN15E6中,构建重组质粒pN15E6-M-lysis、pN15E6-mE和pN15E6-E,分别转化入JM109和DH31soplacI二种大肠杆菌中,观察重组大肠杆菌在含IPTG诱导剂平板培养基上的生长情况,判定M-lysis、mE、E基因过表达对大肠杆菌宿主的影响。结果显示,在大肠杆菌JM109菌株中,mE基因过表达仅表现出抑制细菌生长的效果;M-lysis基因过表达部分致死细菌,存活菌生长受到抑制;E基因过表达能够致死绝大部分细菌,但有少量耐受菌生长。在大肠杆菌DH31soplacI菌株中,M-lysis与mE基因过表达也能致死绝大部分细菌,有少量耐受菌生长;而E基因过表达则完全致死涂布于平板上经106稀释的菌株,未见任何菌落生长。将重组菌株稀释后经平板菌落计数,测得M-lysis、mE、E基因过表达后对DH31soplacI菌株致死率分别为99.03 %、99.68 %和99.9998 %,表明E基因过表达后对宿主菌的致死作用最强;同时定期测定培养的3种重组菌OD600nm值,并绘制生长曲线,结果显示过表达E基因能够迅速致死和裂解细菌。本研究为进一步开发噬菌体裂解肽奠定基础,也为探究细菌致死机理提供了一种新模型。
  
Abstract:
In order to investigate the lethality of three phage lysis genes to host Escherichia coli strains, the bacteriophage lysis genes M-lysis, mE and E were cloned into the highly regulated expression vector pN15E6, respectively, in this study. The lethality of over-expression of M-lysis, mE and E genes to E.coli strains JM109 and DH31soplacI was evaluated through plating. The results indicated that over-expression of mE and M-lysis genes only inhibited bacterial growth in JM109 strain, while over-expression of E gene was lethal to its host bacteria, with a small proportion of resistant colonies grown. In DH31soplacI strain, over-expression of M-lysis and mE genes could also kill bacteria, with a small proportion of resistant colonies grown, while over-expression of E gene killed almost all bacteria plated. Statistically, the lethality of M-lysis, mE and E genes over-expressed in DH31soplacI strain were 99.03%, 99.68% and 99.9998%, further confirming that the E gene over-expression exhibited the highest lethality. Growth curves showed that over-expression of E gene rapidly killed and lysed bacteria after IPTG induction. This study laid a foundation for further development of phage lysis peptides, and also provided a new model for exploring the mechanism of bacteria death.

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(本文编辑:彭永刚;英文编辑:蔡文通)

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更新日期/Last Update: 2020-01-19