A5081187257- Microbial infections and disease research
- Bacteriophages and microbial interactions
- Aquaculture disease management and microbiota
- CRISPR and Genetic Engineering
- Plant Virus Research Studies
- Mycobacterium research and diagnosis
- Genomics and Phylogenetic Studies
- Microbial Metabolic Engineering and Bioproduction
- Phytoplasmas and Hemiptera pathogens
- Fungal and yeast genetics research
- Myxozoan Parasites in Aquatic Species
- RNA and protein synthesis mechanisms
- Insect symbiosis and bacterial influences
- Bacterial Genetics and Biotechnology
- RNA modifications and cancer
- Plant Pathogenic Bacteria Studies
- Herpesvirus Infections and Treatments
- Bacterial Infections and Vaccines
- Cancer-related molecular mechanisms research
- Plant Disease Resistance and Genetics
- Pneumonia and Respiratory Infections
- Evolution and Genetic Dynamics
- Toxoplasma gondii Research Studies
- Metabolism and Genetic Disorders
- Parasitic Infections and Diagnostics
Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement
2018-2025
Biologie du Fruit et Pathologie
2016-2025
Université de Bordeaux
2016-2025
J. Craig Venter Institute
2007-2012
Google (United States)
2012
Institut National de la Recherche Agronomique
2005-2012
Laboratoire Génome et Développement des Plantes
2001-2010
Centre Hospitalier Universitaire de Poitiers
1994
We report the design, synthesis, and assembly of 1.08-mega-base pair Mycoplasma mycoides JCVI-syn1.0 genome starting from digitized sequence information its transplantation into a M. capricolum recipient cell to create new cells that are controlled only by synthetic chromosome. The DNA in is designed sequence, including "watermark" sequences other gene deletions polymorphisms, mutations acquired during building process. have expected phenotypic properties capable continuous self-replication.
As a step toward propagation of synthetic genomes, we completely replaced the genome bacterial cell with one from another species by transplanting whole as naked DNA. Intact genomic DNA Mycoplasma mycoides large colony (LC), virtually free protein, was transplanted into capricolum cells polyethylene glycol–mediated transformation. Cells selected for tetracycline resistance, carried M. LC chromosome, contain complete donor and are detectable recipient sequences. These that result...
We recently reported the chemical synthesis, assembly, and cloning of a bacterial genome in yeast. To produce synthetic cell, must be transferred from yeast to receptive cytoplasm. Here we describe methods accomplish this. cloned Mycoplasma mycoides as centromeric plasmid then transplanted it into capricolum viable M. cell. While yeast, was altered by using genetic systems new strain mycoides. These allow construction strains that could not produced with tools available for this bacterium.
Mycoplasmas are commonly described as the simplest self-replicating organisms, whose evolution was mainly characterized by genome downsizing with a proposed evolutionary scenario similar to that of obligate intracellular bacteria such insect endosymbionts. Thus far, analysis mycoplasma genomes indicates low level horizontal gene transfer (HGT) implying DNA acquisition is strongly limited in these minimal bacteria. In this study, ruminant pathogen Mycoplasma agalactiae sequenced. Comparative...
Most microbes have not been cultured, and many of those that are cultivatable difficult, dangerous or expensive to propagate genetically intractable. Routine cloning large genome fractions whole genomes from these organisms would significantly enhance their discovery genetic functional characterization. Here we report the bacterial in yeast Saccharomyces cerevisiae as single-DNA molecules. We cloned Mycoplasma genitalium (0.6 Mb), M. pneumoniae (0.8 Mb) mycoides subspecies capri (1.1...
In eukaryotes, DNA-associated protein complexes coevolve with genomic sequences to orchestrate chromatin folding. We investigate the relationship between DNA sequence and spontaneous loading activity of components in absence coevolution. Using bacterial genomes integrated into Saccharomyces cerevisiae , which diverged from yeast more than 2 billion years ago, we show that nucleosomes, cohesins, associated transcriptional machinery can lead formation two different archetypes, one transcribed...
Genome transplantation (GT) allows the installation of purified chromosomes into recipient cells, causing resulting organisms to adopt genotype and phenotype conferred by donor cells. This key process remains a bottleneck in synthetic biology, especially for genome engineering strategies intractable economically important microbial species. So far, this has only been reported using two closely related bacteria, Mycoplasma mycoides subsp. capri (Mmc) capricolum (Mcap), main factors driving...
One remarkable achievement in synthetic biology was the reconstruction of mycoplasma genomes and their cloning yeast where they can be modified using available genetic tools. Recently, CRISPR/Cas9 editing tools were developed for mutagenesis. Here, we report adaptation engineering bacterial cloned yeast. A seamless deletion glycerol-3-phosphate oxidase-encoding gene (glpO) achieved without selection one step, 90 nt paired oligonucleotides as templates to drive recombination. Screening...
Development of a new generation vaccines is key challenge for the control infectious diseases affecting both humans and animals. Synthetic biology methods offer ways to engineer bacterial chassis that can be used as vectors present heterologous antigens train immune system against pathogens. Here, we describe construction based on fast-growing Mycoplasma feriruminatoris, first steps toward its application live vaccine contagious caprine pleuropneumonia (CCPP). To do so, M. feriruminatoris...
Recently, artificial oriC plasmids containing the chromosomal dnaA gene and surrounding DnaA box sequences were obtained for mollicutes Spiroplasma citri Mycoplasma pulmonis. In order to study specificity of these among mollicutes, a set similar was developed three mycoplasmas belonging mycoides cluster, subsp. LC (MmmLC), M.mycoides SC (MmmSC) capricolum subsp . Mycoplasmas from S.citri M.pulmonis used as recipients transformation experiments by homologous heterologous plasmids. All five...
Cloning and transplantation of bacterial genomes is a powerful method for the creation engineered microorganisms. However, much remains to be understood about molecular mechanisms limitations this approach. We report whole-genome cloning Mesoplasma florum in Saccharomyces cerevisiae, use model investigate impact chromosome yeast cells. Our results indicate that cloned M. genome subjected weak transcriptional activity, causes no significant on growth. also can transplanted into Mycoplasma...
Efficient protein synthesis in all organisms requires the post-transcriptional methylation of specific ribosomal ribonucleic acid (rRNA) and transfer RNA (tRNA) nucleotides. The reactions are almost invariably catalyzed by enzymes that use S-adenosylmethionine (AdoMet) as methyl group donor. One noteworthy exception is seen some bacteria, where conserved tRNA at m5U54 added enzyme TrmFO using flavin adenine dinucleotide together with N5,N10-methylenetetrahydrofolate one-carbon minimalist...
Over the past decade, a new strategy was developed to bypass difficulties genetically engineer some microbial species by transferring (or "cloning") their genome into another organism that is amenable efficient genetic modifications and therefore acts as living workbench. As such, yeast Saccharomyces cerevisiae has been used clone genomes from viruses, bacteria, algae. The cloning step requires insertion of elements in interest, order drive its replication maintenance an artificial...
Mycoplasmas are the smallest free-living organisms and cause a number of economically important diseases affecting humans, animals, insects plants. Here, we demonstrate that highly virulent Mycoplasma mycoides subspecies capri (Mmc) can be fully attenuated via targeted deletion non-essential genes encoding, among others, potential virulence traits. Five genomic regions, representing approximately ten percent original Mmc genome, were successively deleted using Saccharomyces cerevisiae as an...
ABSTRACT Mycoplasma pulmonis is a natural rodent pathogen, considered privileged model for studying respiratory mycoplasmosis. The complete genome of this bacterium, which belongs to the class Mollicutes , has recently been sequenced, but role specific genes requires improved genetic tools. In silico comparative analysis sequenced mollicute genomes indicated lack conservation gene order in region containing predicted origin replication ( oriC ) and existence, most examined, putative DnaA...
Mycoplasma mycoides subsp. capri (Mmc) and (Mmm) are important ruminant pathogens worldwide causing diseases such as pleuropneumonia, mastitis septicaemia. They express galactofuranose residues on their surface, but role in pathogenesis has not yet been determined. The M. genomes contain up to several copies of the glf gene, which encodes an enzyme catalysing last step synthesis galactofuranose. We generated a deletion gene strain Mmc using genome transplantation tandem repeat endonuclease...
MIB and MIP bind to antibodies disrupt the antigen binding site promote dissociation of antibody-antigen interactions.
ABSTRACT Replicative oriC plasmids were recently developed for several mollicutes, including three Mycoplasma species belonging to the mycoides cluster that are responsible bovine and caprine diseases: subsp. small-colony type, large-colony capricolum . In this study, evaluated in M. as genetic tools (i) expression of heterologous proteins (ii) gene inactivation by homologous recombination. The reporter lacZ , encoding β-galactosidase, spiralin, an abundant surface lipoprotein related...
With the development of several new technologies using synthetic biology, it is possible to engineer genetically intractable organisms including Mycoplasma mycoides subspecies capri (Mmc), by cloning intact bacterial genome in yeast, host yeast's genetic tools modify cloned genome, and subsequently transplanting modified into a recipient cell obtain mutant cells encoded genome. The recently described tandem repeat coupled with endonuclease cleavage (TREC) method has been successfully used...
Clustered regularly interspaced short palindromic repeat (CRISPR)-Cas systems are bacterial defences that target bacteriophages and mobile genetic elements. How these evolve in novel host environments remains largely unknown. We studied the evolution of CRISPR-Cas system
ABSTRACT Over the past several years, significant advances have been made in molecular genetics of Mollicutes (the simplest cells that can be grown axenic culture). Nevertheless, a number basic tools are still required before genetic manipulations become routine. Here we describe development new dominant selectable marker based on enzyme puromycin- N -acetyltransferase from Streptomyces alboniger . Puromycin is an antibiotic mimics 3′-terminal end aminoacylated tRNAs and attaches to carboxyl...
Mycoplasmas are minimal bacteria that infect humans, wildlife, and most economically relevant livestock species. Mycoplasma infections cause a large range of chronic inflammatory diseases, eventually leading to death in some animals. Due the lack efficient recombination genome engineering tools for species, production mutant strains identification virulence factors development improved vaccine is limited. Here, we demonstrate adaptation an Cas9-Base Editor system introduce targeted mutations...
Abstract Bacterial cell shape is generally determined through an interplay between the peptidoglycan wall and cytoplasmic filaments made of polymerized MreB. Indeed, some bacteria (e.g., Mycoplasma ) that lack both a mreB genes consist non-motile cells are spherical or pleomorphic. However, other members same class Mollicutes Spiroplasma , also lacking wall) display helical kink-based motility, which thought to rely on presence five MreB isoforms specific fibril protein. Here, we show...