Abstract:
BACKGROUND:Microbial genome editing is a powerful tool to modify chromosome in way of deletion, insertion or replacement, which is one of the most important techniques in metabolic engineering research. The emergence of CRISPR/Cas9 technique inspires various genomic editing methods. RESULTS:In this research, the goal of development of a fast and easy method for Escherichia coli genome editing with high efficiency is pursued. For this purpose, we designed modular plasmid assembly strategy, compared effects of different length of homologous arms for recombination, and tested different sets of recombinases. The final technique we developed only requires one plasmid construction and one transformation of practice to edit a genomic locus with 3 days and minimal lab work. In addition, the single temperature sensitive plasmid is easy to eliminate for another round of editing. Especially, process of the modularized editing plasmid construction only takes 4 h. CONCLUSION:In this study, we developed a fast and easy genome editing procedure based on CRISPR/Cas9 system that only required the work of one plasmid construction and one transformation, which allowed modification of a chromosome locus within 3 days and could be performed continuously for multiple loci.
journal_name
Microb Cell Factjournal_title
Microbial cell factoriesauthors
Zhao D,Yuan S,Xiong B,Sun H,Ye L,Li J,Zhang X,Bi Cdoi
10.1186/s12934-016-0605-5subject
Has Abstractpub_date
2016-12-01 00:00:00pages
205issue
1issn
1475-2859pii
10.1186/s12934-016-0605-5journal_volume
15pub_type
杂志文章abstract:BACKGROUND:Mucosal delivery of therapeutic proteins using genetically modified strains of lactic acid bacteria (gmLAB) is being investigated as a new therapeutic strategy. METHODS:We developed a strain of gmLAB, Lactococcus lactis NZ9000 (NZ-HO), which secretes the anti-inflammatory molecule recombinant mouse heme oxy...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0378-2
更新日期:2015-11-25 00:00:00
abstract:BACKGROUND:Chinese Nong-flavor (NF) liquor is continuously and stably produced by solid-state fermentation technology for 1000 years, resulting in enrichment of special microbial community and enzymes system in its starter. Based on traditional culture-dependent methods, these functional enzymes are hardly obtained. Ac...
journal_title:Microbial cell factories
pub_type: 杂志文章
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-10-15
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-12-1
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-014-0145-9
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
pub_type: 杂志文章
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journal_title:Microbial cell factories
pub_type: 杂志文章
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-016-0527-2
更新日期:2016-07-28 00:00:00
abstract::Nanotechnology is increasingly using both materials and nano-objects synthesized by living beings, most of them produced by microbial cells. Emerging technologies and highly integrative approaches (such as 'omics and systems biology), that have been largely proven successful for the production of proteins and secondar...
journal_title:Microbial cell factories
pub_type: 社论
doi:10.1186/1475-2859-9-53
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-018-0897-8
更新日期:2018-03-26 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-015-0309-2
更新日期:2015-08-12 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-6-27
更新日期:2007-08-20 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01369-2
更新日期:2020-06-11 00:00:00
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journal_title:Microbial cell factories
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doi:10.1186/s12934-019-1158-1
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-020-01308-1
更新日期:2020-02-19 00:00:00
abstract::BACKGROUND: A rapid microtiter plate based sandwich hybridization assay was developed for detection and quantification of single RNA species using magnetic beads. Following solution hybridization target RNA molecules were collected by biotin-streptavidin affinity binding and detected by fluorescence signal generated b...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-2-4
更新日期:2003-04-28 00:00:00
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journal_title:Microbial cell factories
pub_type: 杂志文章
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journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/s12934-017-0797-3
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abstract:BACKGROUND:E. coli is a robust host for various genetic manipulations and has been used commonly for bioconversion of hexose and pentose sugars into valuable products. One of the products that E. coli make under fermentative condition is ethanol. However, availability of limited reducing equivalence and generation of c...
journal_title:Microbial cell factories
pub_type: 杂志文章
doi:10.1186/1475-2859-11-145
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journal_title:Microbial cell factories
pub_type: 杂志文章
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journal_title:Microbial cell factories
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journal_title:Microbial cell factories
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