相关实验视频

Updated: Sep 10, 2025

Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis

Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis

Published on: October 31, 2014

工程Zymomonas mobilis用于改善多基因生物合成途径的基因转化和稳定性

Yuhuan Huang¹, Xiaojie Wang², Mao Chen³

¹Biomass Energy Technology Research Centre, Key Laboratory of Development and Application of Rural Renewable Energy (Ministry of Agriculture and Rural Affairs), Biogas Institute of Ministry of Agriculture and Rural Affairs, Chengdu 610041, China; Graduate School of Chinese Academy of Agricultural Science, Beijing 100081, China.

Bioresource technology

|August 24, 2025

概括

这项研究通过增强等离子体转化和稳定性来改进Zymomonas mobilis的代谢工程. 关键基因的淘汰导致三重缺陷突变产生显著更多的β-胡卜素.

科学领域:

代谢工程
合成生物学
微生物生物技术

背景情况:

这种微生物对新陈代谢工程具有前景.
目前的局限性包括低效的基因转化和等离子体不稳定性,特别是大于8kB的等离子体.

研究的目的:

开发对Zymomonas mobilis进行改进的遗传工具.
提高Z. mobilis的转化效率和等离子体稳定性.
在Z. mobilis中增加β- 胡卜素的产生.

主要方法:

为Z. mobilis ZM4建立一个促进器和终结器库.
在pEZ-crt1和pEZ-crt2等离子体上设计β-胡卜素基因集群 (crtEXYIB).
参与限制修改 (R-M) 和DNA修复系统 (mrr,hsdM,tatD) 的基因的识别和淘汰.

主要成果:

已确定用于生产β-胡卜素的高产品种CRT1-29 (0.93 mg/g DCW) 和CRT2-22 (1.25 mg/g DCW).
mrr和hsdM的淘汰显著提高了甲基化等离子体的转化效率,并使9.5kb等离子体的电穿孔成为可能.
删除tatD改善了外源性等离子体的稳定性,三重突变MHT达到2. 09 mg/ g的β- 胡卜素DCW.

结论:

开发的调控元素和基因改造克服了Z. mobilis工程中的关键障碍.
三重缺陷突变MHT表现出增强的转化和稳定性,可产生高水平的β- 胡卜素.
这项工作促进了Z. mobilis在代谢工程和合成生物学中的更广泛应用.

关键词:

修复DNA的基因基因稳定性基因转化金门组装限制修改系统

更多相关视频

Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX

Inducible T7 RNA Polymerase-mediated Multigene Expression System, pMGX

Published on: June 27, 2017

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Rapid Assembly of Multi-Gene Constructs using Modular Golden Gate Cloning

Published on: February 5, 2021

相关概念视频

Transformation

Transformation

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...

Transgenic Plants

Transgenic Plants

Recombinant DNA technology called transgenesis is often used to add a foreign gene or remove a detrimental gene from an organism. Such genetically modified organisms are called transgenic organisms.
The first-ever transgenic plant was a tobacco plant developed in 1983 that showed resistance against the tobacco mosaic virus. Since then, many transgenic plants have been developed and commercialized for improving the agricultural, ornamental, and horticultural value of a crop plant. Transgenic...