ArticleNature protocols2011
Knockout and pullout recombineering for naturally transformable Burkholderia thailandensis and Burkholderia pseudomallei.
Article in Nature protocols, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 26 papers.
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Who cites it
26 citing papers in PubMed.
- CnRed: Efficient, Marker-free Genome Engineering ofACS synthetic biology · 2025Article
- Half a century after their discovery: Structural insights into exonuclease and annealase proteins catalyzing recombineering.Engineering microbiology · 2024Review
- TetR-like regulator BP1026B_II1561 controls aromatic amino acid biosynthesis and intracellular pathogenesis inFrontiers in microbiology · 2024Article
- The emerging role of recombineering in microbiology.Engineering microbiology · 2023Review
- Recombineering enables genome mining of novel siderophores in a non-modelEngineering microbiology · 2023Article
- CRISPR-Associated Transposase for Targeted Mutagenesis in Diverse Proteobacteria.ACS synthetic biology · 2023Article
- Recombineering in Non-Model Bacteria.Current protocols · 2022Article
- Biotechnology approaches for natural product discovery, engineering, and production based on Burkholderia bacteria.Current opinion in biotechnology · 2022Review
- Engineering ofFrontiers in microbiology · 2022Article
- Knockout of a highly GC-rich gene in Burkholderia pyrrocinia by recombineering with freeze-thawing transformation.Molecular plant pathology · 2021Article
- Development and application of an efficient recombineering system for Burkholderia glumae and Burkholderia plantarii.Microbial biotechnology · 2021Article
- Identification of a PadR-type regulator essential for intracellular pathogenesis of Burkholderia pseudomallei.Scientific reports · 2021Article
- The Burkholderia pseudomallei intracellular 'TRANSITome'.Nature communications · 2021Article
- Genomics-Driven Activation of Silent Biosynthetic Gene Clusters inMolecules (Basel, Switzerland) · 2021Article
- Article
- Establishment of recombineering genome editing system in Paraburkholderia megapolitana empowers activation of silent biosynthetic gene clusters.Microbial biotechnology · 2020Article
- Lipid A Remodeling Is a Pathoadaptive Mechanism That Impacts Lipopolysaccharide Recognition and Intracellular Survival of Burkholderia pseudomallei.Infection and immunity · 2018Article
- The heritable natural competency trait of Burkholderia pseudomallei in other Burkholderia species through comE and crp.Scientific reports · 2018Article
- Discovery of recombinases enables genome mining of cryptic biosynthetic gene clusters in Burkholderiales species.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
- An avirulent Burkholderia pseudomallei ∆purM strain with atypical type B LPS: expansion of the toolkit for biosafe studies of melioidosis.BMC microbiology · 2017Article
Corrections and comments
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Authors and funding
6 authors.
Funding
Abstract
Phage λ-Red proteins are powerful tools for pulling and knocking out chromosomal fragments but have been limited to the γ-proteobacteria. Procedures are described here to easily knock out (KO) and pull out (PO) chromosomal DNA fragments from naturally transformable Burkholderia thailandensis and Burkholderia pseudomallei. This system takes advantage of published compliant counterselectable and selectable markers (sacB, pheS, gat and the arabinose-utilization operon) and λ-Red mutant proteins. pheS-gat (KO) or oriT-ColE1ori-gat-ori1600-rep (PO) PCR fragments are generated with flanking 40- to 45-bp homologies to targeted regions incorporated on PCR primers. One-step recombination is achieved by incubation of the PCR product with cells expressing λ-Red proteins and subsequent selection on glyphosate-containing medium. This procedure takes ~10 d and is advantageous over previously published protocols: (i) smaller PCR products reduce primer numbers and amplification steps, (ii) PO fragments suitable for downstream manipulation in Escherichia coli are obtained and (iii) chromosomal KO increases flexibility for downstream processing.
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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.