ArticleInfection and immunity2006
Genome engineering in Bacillus anthracis using Cre recombinase.
Article in Infection and immunity, 2006. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 54 papers.
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Who cites it
54 citing papers in PubMed, 78 citations in OpenAlex.
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- S9.6-based hybrid capture immunoassay for pathogen detection.Scientific reports · 2023Article
- Development and application of CRISPR-based genetic tools in Bacillus species and Bacillus phages.Journal of applied microbiology · 2022Review
- ClpC-Mediated Sporulation Regulation at Engulfment Stage inIndian journal of microbiology · 2021Article
- Metabolic differentiation and intercellular nurturing underpin bacterial endospore formation.Science advances · 2021Article
- Frontline Science: Anthrax lethal toxin-induced, NLRP1-mediated IL-1β release is a neutrophil and PAD4-dependent event.Journal of leukocyte biology · 2020Article
- Highly Efficient Genome Engineering inFrontiers in microbiology · 2019Article
- Development and evaluation of an efficient heterologous gene knock-in reporter system in Lactococcus lactis.Microbial cell factories · 2017Article
- Article
- TheFrontiers in cellular and infection microbiology · 2017Article
- A Diverse Set of Single-domain Antibodies (VHHs) against the Anthrax Toxin Lethal and Edema Factors Provides a Basis for Construction of a Bispecific Agent That Protects against Anthrax Infection.The Journal of biological chemistry · 2016Article
- Mob/oriT, a mobilizable site-specific recombination system for unmarked genetic manipulation in Bacillus thuringiensis and Bacillus cereus.Microbial cell factories · 2016Article
- Adenoviral Expression of a Bispecific VHH-Based Neutralizing Agent That Targets Protective Antigen Provides Prophylactic Protection from Anthrax in Mice.Clinical and vaccine immunology : CVI · 2016Article
- Sulforaphane inhibits multiple inflammasomes through an Nrf2-independent mechanism.Journal of leukocyte biology · 2016Article
- Bacillus anthracis Overcomes an Amino Acid Auxotrophy by Cleaving Host Serum Proteins.Journal of bacteriology · 2015Article
- The cyclopentenone prostaglandin 15d-PGJ2 inhibits the NLRP1 and NLRP3 inflammasomes.Journal of immunology (Baltimore, Md. : 1950) · 2015Article
- A heterodimer of a VHH (variable domains of camelid heavy chain-only) antibody that inhibits anthrax toxin cell binding linked to a VHH antibody that blocks oligomer formation is highly protective in an anthrax spore challenge model.The Journal of biological chemistry · 2015Article
- clpC operon regulates cell architecture and sporulation in Bacillus anthracis.Environmental microbiology · 2015Article
- Identification of three noncontiguous regions on Bacillus anthracis plasmid pXO1 that are important for its maintenance.Journal of bacteriology · 2014Article
- Rapid targeted gene disruption in Bacillus anthracis.BMC biotechnology · 2013Article
Corrections and comments
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Authors and funding
5 authors at 2 institutions in 1 country.
Funding
Abstract
Genome engineering is a powerful method for the study of bacterial virulence. With the availability of the complete genomic sequence of Bacillus anthracis, it is now possible to inactivate or delete selected genes of interest. However, many current methods for disrupting or deleting more than one gene require use of multiple antibiotic resistance determinants. In this report we used an approach that temporarily inserts an antibiotic resistance marker into a selected region of the genome and subsequently removes it, leaving the target region (a single gene or a larger genomic segment) permanently mutated. For this purpose, a spectinomycin resistance cassette flanked by bacteriophage P1 loxP sites oriented as direct repeats was inserted within a selected gene. After identification of strains having the spectinomycin cassette inserted by a double-crossover event, a thermo-sensitive plasmid expressing Cre recombinase was introduced at the permissive temperature. Cre recombinase action at the loxP sites excised the spectinomycin marker, leaving a single loxP site within the targeted gene or genomic segment. The Cre-expressing plasmid was then removed by growth at the restrictive temperature. The procedure could then be repeated to mutate additional genes. In this way, we sequentially mutated two pairs of genes: pepM and spo0A, and mcrB and mrr. Furthermore, loxP sites introduced at distant genes could be recombined by Cre recombinase to cause deletion of large intervening regions. In this way, we deleted the capBCAD region of the pXO2 plasmid and the entire 30 kb of chromosomal DNA between the mcrB and mrr genes, and in the latter case we found that the 32 intervening open reading frames were not essential to growth.
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Registered trials
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.