相关实验视频
Updated: May 7, 2026
11:57
Electroporation of Mycobacteria
Published on: May 23, 2008
25.3K
使用电穿孔进行Bifidobacterium animalis AR668-R1的高效转化
Yaping Liu1, Zhiqiang Xiong1, Wenhao Liu1
1Shanghai Engineering Research Center of Food Microbiology, School of Health Science and Engineering, University of Shanghai for Science and Technology, Shanghai 200093, China.
Journal of biotechnology
|April 17, 2025
概括
优化电穿孔参数显著提高了Bifidobacterium animalis AR668-R1的转化效率,提高了79倍. 这种改进与高调的能力基因,特别是comEC有关,这促进了基因操纵.
科学领域:
- 微生物学 微生物学
- 分子生物学分子生物学
- 基因工程是一种基因工程.
背景情况:
- 生物菌 (Bifidobacterium animalis) 是一种普遍存在的肠道细菌,用于乳制品.
- 耐氧菌株AR668-R1由于电穿孔效率低,对分子研究提出了挑战.
- 有效的基因转换对于理解和操纵B. animalis. 非常重要.
研究的目的:
- 为了在B. animalis AR668-R1.1.中实现可复制和高的电解介导转换效率.
- 调查转化效率改进背后的分子机制.
- 为了促进B. animalis的基因操纵和功能分析.
主要方法:
- 系统优化电穿孔参数,包括注射剂,砂糖,NaCl,生长阶段,等离子体度,电场和复苏时间.
- 转录分析以确定优化后差异表达的基因.
- 基因过度表达和关键能力基因的淘汰实验 (ssb,基因0596,comEC,基因1115).
主要成果:
- 在优化条件下,转换效率提高了79倍 (3.97 × 10^5 CFU/μg DNA).
- 在优化后,能力基因 (ssb,基因0596,comEC,基因1115) 显著上调.
- 过度表达comEC使转换效率提高了2.5倍;comEC淘汰将其降低了74.9倍.
结论:
- 通过参数优化,可以显著提高B. animalis AR668-R1的电穿孔效率.
- 能力基因的升级和功能,特别是comEC,对于转换效率至关重要.
- 优化的协议和已识别的基因使得B. animalis. 的先进遗传研究成为可能.
相关概念视频
01:33Bacterial Transformation
In 1928, bacteriologist Frederick Griffith worked on a vaccine for pneumonia, which is caused by Streptococcus pneumoniae bacteria. Griffith studied two pneumonia strains in mice: one pathogenic and one non-pathogenic. Only the pathogenic strain killed host mice.
Griffith made an unexpected discovery when he killed the pathogenic strain and mixed its remains with the live, non-pathogenic strain. Not only did the mixture kill host mice, but it also contained living pathogenic bacteria that...
Griffith made an unexpected discovery when he killed the pathogenic strain and mixed its remains with the live, non-pathogenic strain. Not only did the mixture kill host mice, but it also contained living pathogenic bacteria that...
01:33Bacterial Transformation
In 1928, bacteriologist Frederick Griffith worked on a vaccine for pneumonia, which is caused by Streptococcus pneumoniae bacteria. Griffith studied two pneumonia strains in mice: one pathogenic and one non-pathogenic. Only the pathogenic strain killed host mice.
Griffith made an unexpected discovery when he killed the pathogenic strain and mixed its remains with the live, non-pathogenic strain. Not only did the mixture kill host mice, but it also contained living pathogenic bacteria that...
Griffith made an unexpected discovery when he killed the pathogenic strain and mixed its remains with the live, non-pathogenic strain. Not only did the mixture kill host mice, but it also contained living pathogenic bacteria that...
01:26Transformation
Microbial communities are dynamic environments where cell lysis releases free DNA into the surroundings. Other cells can take up this extracellular DNA through a process known as transformation.When a cell incorporates this foreign DNA into its genome, resulting in genetic modification, the process is known as transformation. Cells capable of this process are termed competent. Competence can be natural, as observed in certain bacteria and archaea, or artificially induced in the...