Large‐scale transposon mutagenesis of<i>Mycoplasma pulmonis</i>

French, Christopher T.; Lao, Ping; Loraine, Ann E.; Matthews, Brian T.; Yu, Huilan; Dybvig, Kevin

doi:10.1111/j.1365-2958.2008.06262.x

Cited by 113 publications

(121 citation statements)

References 37 publications

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“…Several genome-scale identifications of such genes have been performed in prokaryotic and eukaryotic organisms by different groups with different strategies. Generally, there are three main experimental approaches to identify essential genes under particular growth conditions: massive transposon mutagenesis strategies (Judson and Mekalanos, 2000;Akerley et al, 2002;Salama et al, 2004;French et al, 2008), the use of antisense RNA (Ji et al, 2001;Forsyth et al, 2002) to inhibit gene expression, and the systematic inactivation of each individual gene present in a genome (Herring et al, 2003;Kobayashi et al, 2003). However, these results showed that percentage of essential genes in the whole genome is really low (Table 2).…”

Section: Experimental Gene Inactivation Approachmentioning

confidence: 39%

How to make a minimal genome for synthetic minimal cell

2010

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As a key focus of synthetic biology, building a minimal artificial cell has given rise to many discussions. A synthetic minimal cell will provide an appropriate chassis to integrate functional synthetic parts, devices and systems with functions that cannot generally be found in nature. The design and construction of a functional minimal genome is a key step while building such a cell/ chassis since all the cell functions can be traced back to the genome. Kinds of approaches, based on bioinformatics and molecular biology, have been developed and proceeded to derive essential genes and minimal gene sets for the synthetic minimal genome. Experiments about streamlining genomes of model bacteria revealed genome reduction led to unanticipated beneficial properties, such as high electroporation efficiency and accurate propagation of recombinant genes and plasmids that were unstable in other strains. Recent achievements in chemical synthesis technology for large DNA segments together with the rapid development of the wholegenome sequencing, have transferred synthesis of genes to assembly of the whole genomes based on oligonucleotides, and thus created strong preconditions for synthesis of artificial minimal genome. Here in this article, we review briefly the history and current state of research in this field and summarize the main methods for making a minimal genome. We also discuss the impacts of minimized genome on metabolism and regulation of artificial cell.

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Section: Experimental Gene Inactivation Approachmentioning

confidence: 39%

How to make a minimal genome for synthetic minimal cell

2010

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“…Cyclic di-AMP secreted into the cytosol of host cells by multiefflux pumps of the intracellular pathogen L. monocytogenes activates a cytosolic surveillance pathway in host immune cells such as macrophages (447). This potential role of c-di-AMP in virulence and stress resistance, in combination with the fact that genes coding for diadenylate cyclases are essential in intracellular pathogens (451)(452)(453)(454), makes c-di-AMP signaling pathways an attractive target for antimicrobial therapy.…”

Section: The Novel Cyclic Dinucleotide Second Messengers Cyclic Di-ammentioning

confidence: 99%

Cyclic di-GMP: the First 25 Years of a Universal Bacterial Second Messenger

Römling

Galperin

Gomelsky

2013

Microbiol Mol Biol Rev

1,471

1,698

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SUMMARY Twenty-five years have passed since the discovery of cyclic dimeric (3′→5′) GMP (cyclic di-GMP or c-di-GMP). From the relative obscurity of an allosteric activator of a bacterial cellulose synthase, c-di-GMP has emerged as one of the most common and important bacterial second messengers. Cyclic di-GMP has been shown to regulate biofilm formation, motility, virulence, the cell cycle, differentiation, and other processes. Most c-di-GMP-dependent signaling pathways control the ability of bacteria to interact with abiotic surfaces or with other bacterial and eukaryotic cells. Cyclic di-GMP plays key roles in lifestyle changes of many bacteria, including transition from the motile to the sessile state, which aids in the establishment of multicellular biofilm communities, and from the virulent state in acute infections to the less virulent but more resilient state characteristic of chronic infectious diseases. From a practical standpoint, modulating c-di-GMP signaling pathways in bacteria could represent a new way of controlling formation and dispersal of biofilms in medical and industrial settings. Cyclic di-GMP participates in interkingdom signaling. It is recognized by mammalian immune systems as a uniquely bacterial molecule and therefore is considered a promising vaccine adjuvant. The purpose of this review is not to overview the whole body of data in the burgeoning field of c-di-GMP-dependent signaling. Instead, we provide a historic perspective on the development of the field, emphasize common trends, and illustrate them with the best available examples. We also identify unresolved questions and highlight new directions in c-di-GMP research that will give us a deeper understanding of this truly universal bacterial second messenger.

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“…The 2 DGA and RHR motifs that are conserved in DacA proteins are also conserved in ssDacA. DacA orthologs appear to be essential for the viability of bacteria, as they cannot be deleted using traditional genetic techniques (Song et al, 2005;Glass et al, 2006;French et al, 2008;Woodward et al, 2010;Corrigan et al, 2011), and the significance of c-di-AMP in reported pathogens imply an important role of this protein in SS2 pathogenesis. In this study, we report this functional diadenylate cyclase (ssDacA) in SS2.…”

Section: Introductionmentioning

confidence: 99%

Diadenylate cyclase evaluation of ssDacA (SSU98_1483) in Streptococcus suis serotype 2

Du¹,

Sun²

2015

Genet. Mol. Res.

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ABSTRACT. Cyclic diadenosine monophosphate is a recently identified signaling molecule. It has been shown to play important roles in bacterial pathogenesis. SSU98_1483 (ssDacA), which is an ortholog of Listeria monocytogenes DacA, is a putative diadenylate cyclase in Streptococcus suis serotype 2. In this study, we determined the enzymatic activity of ssDacA in vitro using high-performance liquid chromatography and mass spectrometry. Our results showed that ssDacA was a diadenylate cyclase that converts ATP into cyclic diadenosine monophosphate in vitro. The diadenylate cyclase activity of ssDacA was dependent on divalent metal ions such as Mg , and it is more active under basic pH than under acidic pH. The conserved RHR motif in ssDacA was essential for its enzymatic activity, and mutation in this motif abolished the diadenylate cyclase activity of ssDacA. These results indicate that ssDacA is a diadenylate cyclase, which synthesizes cyclic diadenosine monophosphate in Streptococcus suis serotype 2.

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Large‐scale transposon mutagenesis ofMycoplasma pulmonis

Cited by 113 publications

References 37 publications

How to make a minimal genome for synthetic minimal cell

How to make a minimal genome for synthetic minimal cell

Cyclic di-GMP: the First 25 Years of a Universal Bacterial Second Messenger

Diadenylate cyclase evaluation of ssDacA (SSU98_1483) in Streptococcus suis serotype 2

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