• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

优化电穿孔条件以将异源 DNA 导入.

Optimization of Electroporation Conditions for Introducing Heterologous DNA into .

机构信息

National Biotechnology Policy Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Republic of Korea.

School of Earth Sciences and Environmental Engineering, Gwangju Institute of Science and Technology (GIST), Gwangju 61005, Republic of Korea.

出版信息

J Microbiol Biotechnol. 2024 Nov 28;34(11):2347-2352. doi: 10.4014/jmb.2408.08044. Epub 2024 Sep 20.

DOI:10.4014/jmb.2408.08044
PMID:39403725
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11637821/
Abstract

is a strain capable of both photoautotrophic and chemoautotrophic growth, with various metabolic pathways that make it highly suitable for converting carbon dioxide into high value-added products. However, its low transformation efficiency has posed challenges for genetic and metabolic engineering of this strain. In this study, we aimed to increase the transformation efficiency of by deleting the gene coding for an endogenous DNA restriction enzyme that inhibits. We evaluated the effects of growth conditions for making electrocompetent cells and optimized electroporation parameters to be a cuvette width of 0.1 cm, an electric field strength of 30 kV/cm, a resistance of 200 Ω, and a plasmid DNA amount of 0.5 μg, followed by a 24-h recovery period. As a result, we observed over 7,000 transformants per μg of DNA under the optimized electroporation conditions using the strain, which is approximately 10 times higher than that of wild-type under standard bacterial electroporation conditions. These findings are expected to enhance the application of in various industrial fields in the future.

摘要

是一种既能进行光合作用又能进行化能自养生长的菌株，具有多种代谢途径，使其非常适合将二氧化碳转化为高附加值产品。然而，其低转化效率给该菌株的遗传和代谢工程带来了挑战。在这项研究中，我们旨在通过删除编码内源性 DNA 限制酶的基因来提高 的转化效率，该酶抑制 。我们评估了用于制备电感受态细胞的生长条件，并优化了电穿孔参数，电穿孔参数为：小室宽度为 0.1 厘米，电场强度为 30 kV/cm，电阻为 200 Ω，质粒 DNA 量为 0.5 μg，随后进行 24 小时恢复。结果，我们在优化的电穿孔条件下使用 菌株观察到每微克 DNA 的转化子超过 7000 个，这大约是标准细菌电穿孔条件下野生型 的 10 倍。这些发现有望提高 在未来各种工业领域的应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6aa4/11637821/8d50464af60a/jmb-34-11-2347-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6aa4/11637821/5d01eab3a5a0/jmb-34-11-2347-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6aa4/11637821/fb2c0ce4493c/jmb-34-11-2347-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6aa4/11637821/3a29fcc3100c/jmb-34-11-2347-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6aa4/11637821/8d50464af60a/jmb-34-11-2347-f4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6aa4/11637821/5d01eab3a5a0/jmb-34-11-2347-f1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6aa4/11637821/fb2c0ce4493c/jmb-34-11-2347-f2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6aa4/11637821/3a29fcc3100c/jmb-34-11-2347-f3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6aa4/11637821/8d50464af60a/jmb-34-11-2347-f4.jpg

相似文献

1
Optimization of Electroporation Conditions for Introducing Heterologous DNA into .优化电穿孔条件以将异源 DNA 导入.
J Microbiol Biotechnol. 2024 Nov 28;34(11):2347-2352. doi: 10.4014/jmb.2408.08044. Epub 2024 Sep 20.
2
Efficient Cas9-based genome editing of Rhodobacter sphaeroides for metabolic engineering.高效基于 Cas9 的红杆菌基因组编辑用于代谢工程。
Microb Cell Fact. 2019 Nov 25;18(1):204. doi: 10.1186/s12934-019-1255-1.
3
Isolation of plasmid DNA sequences that complement Rhodobacter sphaeroides mutants deficient in the capacity for CO2-dependent growth.分离能够互补球形红细菌中二氧化碳依赖生长能力缺陷突变体的质粒DNA序列。
J Gen Microbiol. 1989 Jun;135(6):1699-713. doi: 10.1099/00221287-135-6-1699.
4
Genetic transformation of Rhodopseudomonas sphaeroides by plasmid DNA.球形红假单胞菌的质粒DNA遗传转化
J Bacteriol. 1982 Oct;152(1):89-97. doi: 10.1128/jb.152.1.89-97.1982.
5
Tackling codon usage bias for heterologous expression in Rhodobacter sphaeroides by supplementation of rare tRNAs.通过补充稀有tRNA解决球形红细菌异源表达中的密码子使用偏好问题。
Enzyme Microb Technol. 2015 May;72:25-34. doi: 10.1016/j.enzmictec.2015.02.003. Epub 2015 Feb 16.
6
Differential expression of the CO2 fixation operons of Rhodobacter sphaeroides by the Prr/Reg two-component system during chemoautotrophic growth.球形红细菌的二氧化碳固定操纵子在化学自养生长过程中通过Prr/Reg双组分系统的差异表达。
J Bacteriol. 2002 Dec;184(23):6654-64. doi: 10.1128/JB.184.23.6654-6664.2002.
7
Introduction of the Menaquinone Biosynthetic Pathway into and Synthesis of Menaquinone for Incorporation into Heterologously Expressed Integral Membrane Proteins.导入menaquinone 生物合成途径和合成menaquinone 以整合到异源表达的整合膜蛋白中。
ACS Synth Biol. 2020 May 15;9(5):1190-1200. doi: 10.1021/acssynbio.0c00066. Epub 2020 Apr 21.
8
The optimization system for preparation of TG1 competent cells and electrotransformation.TG1 感受态细胞的制备和电转化优化系统。
Microbiologyopen. 2020 Jul;9(7):e1043. doi: 10.1002/mbo3.1043. Epub 2020 May 11.
9
Use of new strains of Rhodobacter sphaeroides and a modified simple culture medium to increase yield and facilitate purification of the reaction centre.利用新型球形红杆菌菌株和改良的简单培养基来提高反应中心的产量并便于其纯化。
Photosynth Res. 2014 May;120(1-2):197-205. doi: 10.1007/s11120-013-9866-6. Epub 2013 Jun 14.
10
A rapid and efficient electroporation method for transformation of Halomonas sp. O-1.一种用于盐单胞菌属O-1转化的快速高效电穿孔方法。
J Microbiol Methods. 2016 Oct;129:127-132. doi: 10.1016/j.mimet.2016.08.009. Epub 2016 Aug 16.

本文引用的文献

1
Optimization of electrotransformation (ETF) conditions in lactic acid bacteria (LAB).优化乳酸菌（LAB）电转化（ETF）条件。
J Microbiol Methods. 2020 Jul;174:105944. doi: 10.1016/j.mimet.2020.105944. Epub 2020 May 15.
2
Efficient Cas9-based genome editing of Rhodobacter sphaeroides for metabolic engineering.高效基于 Cas9 的红杆菌基因组编辑用于代谢工程。
Microb Cell Fact. 2019 Nov 25;18(1):204. doi: 10.1186/s12934-019-1255-1.
3
Metabolic Redesign of Rhodobacter sphaeroides for Lycopene Production.利用红假单胞菌进行番茄红素的代谢重设计。
J Agric Food Chem. 2018 Jun 13;66(23):5879-5885. doi: 10.1021/acs.jafc.8b00855. Epub 2018 Jun 4.
4
An Efficient Transformation Method for the Bioplastic-Producing "Knallgas" Bacterium Ralstonia eutropha H16.高效转化生产生物塑料的“爆炸气体”假单胞菌 Ralstonia eutropha H16 的方法。
Biotechnol J. 2017 Nov;12(11). doi: 10.1002/biot.201700081. Epub 2017 Sep 5.
5
Metabolic engineering in chemolithoautotrophic hosts for the production of fuels and chemicals.利用化能自养宿主进行代谢工程以生产燃料和化学品。
Metab Eng. 2015 Jul;30:105-120. doi: 10.1016/j.ymben.2015.04.008. Epub 2015 May 8.
6
Restriction-Modification Systems as a Barrier for Genetic Manipulation of Staphylococcus aureus.限制修饰系统作为金黄色葡萄球菌基因操作的障碍
Methods Mol Biol. 2016;1373:9-23. doi: 10.1007/7651_2014_180.
7
Study on the construction of recombined plasmid pMG36e-lacc1 and the electroporation of Lactobacillus buchneri.重组质粒pMG36e-lacc1的构建及布氏乳杆菌电穿孔的研究
Biomed Mater Eng. 2014;24(6):3855-61. doi: 10.3233/BME-141216.
8
A critical combination of operating parameters can significantly increase the electrotransformation efficiency of a gram-positive Dietzia strain.关键的操作参数组合可显著提高革兰氏阳性Dietzia菌株的电转化效率。
J Microbiol Methods. 2014 Aug;103:144-51. doi: 10.1016/j.mimet.2014.05.015. Epub 2014 Jun 2.
9
Optimization of a plasmid electroporation protocol for Aeromonas salmonicida subsp. salmonicida.杀鲑气单胞菌杀鲑亚种质粒电穿孔方案的优化
J Microbiol Methods. 2014 Mar;98:44-9. doi: 10.1016/j.mimet.2013.12.019. Epub 2013 Dec 31.
10
Use of new strains of Rhodobacter sphaeroides and a modified simple culture medium to increase yield and facilitate purification of the reaction centre.利用新型球形红杆菌菌株和改良的简单培养基来提高反应中心的产量并便于其纯化。
Photosynth Res. 2014 May;120(1-2):197-205. doi: 10.1007/s11120-013-9866-6. Epub 2013 Jun 14.