Team:Evry/Biology/Chassis/Pseudovibrio

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<h2> <i>Pseudovibrio denitrificans </i> the bacterium to modify.</h2>
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<h2> <i>Pseudovibrio denitrificans </i> the bacterium to modify</h2>
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== <b><big>From a poorly known genius, a jewel emerged</big></b> ==
== <b><big>From a poorly known genius, a jewel emerged</big></b> ==
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With only 12 species in the genius, and 2 strains sequenced (Pseudovibrio sp. FO-BEG1, Pseudovibrio sp. JE062) we knew we would have to sequence our strain of <i>Pseudovibrio denitrificans </i> with a mapping to one of the 2 strains references. Eventhough in the first place the species was chosen because of its natural denitrification ability (cf <b><big> toxic compound</big></b>) and sourced at <b><big>DMSZ</big></b>.
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At first glance, working on the microbiome of sponges appears like a hazardous task: 5734 articles for marine sponges, only 51 with microbiome of sponges, only 28 articles mentioning <i>Pseudovibrio genius</i>, only 5 mentioning <i>Pseudovibrio denitrificans</i>, and finally none about a genetic engineering system in it.
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In addition to that <i>Pseudovibrio denitrificans </i> or strains related to the species has been shown as majoritary in at least six microbiomes of sponges in the mediterrean sea, where spongia officinalis reside.
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Moreover with only 12 species in the genius, and 2 strains sequenced (Pseudovibrio sp. FO-BEG1, Pseudovibrio sp. JE062) we knew we would have to sequence our strain of <i>Pseudovibrio denitrificans </i> with a mapping to one of the 2 strains references. A formidable drawback even considering that in the first place the species was chosen because of its natural denitrification ability (see <a href="https://2014.igem.org/Team:Evry/Biology/ToxicCompounds"> <b><big> toxic compound</big></b></a>) and easy to source at <a href="http://www.dsmz.de/catalogues/details/culture/DSM-17465.html"> <b><big>DMSZ</big></b></a>.
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On the other hand <i>Pseudovibrio denitrificans</i>, or strains related to the species, has been shown to be in the majority in at least six microbiomes of sponges in the Mediterranean Sea, where <i>spongia officinalis</i> resides.
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Now the thorny question is: can it be easily used  in our lab?
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== <b><big>A bacterium we  could work with</big></b> ==
== <b><big>A bacterium we  could work with</big></b> ==
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Stained as a Gram-negative, the cells in late exponential to early stationary phase of growth were predominantly straight or curved rods. They were motile by means of one to several lateral or subpolar flagella. Both strains required NaCl for growth and exhibited optimal growth at about 30 degrees C, pH 8 and 3 % NaCl.  
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In the literature <i>Pseudovibrio denitrificans </i> is described as a Gram-negative, motile by means of one to several lateral or subpolar flagella requiring NaCl for growth. It exhibits optimal growth at about 30 , pH 8 and 3 % NaCl, and a  doubling time of 45 min in rich media. In a nutshell the conditions seem favourable for an iGEM project</i>.
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They are known to be capable of anaerobic growth by carrying out denitrifying metabolism using nitrate, nitrite or nitrous oxide as terminal electron acceptors. Being consequently a main reason for our <b><big>RNAseq</big></b> study to discover the transcripts upregulated by the presence of nitrite and clone their promoters as new sensors.
They are known to be capable of anaerobic growth by carrying out denitrifying metabolism using nitrate, nitrite or nitrous oxide as terminal electron acceptors. Being consequently a main reason for our <b><big>RNAseq</big></b> study to discover the transcripts upregulated by the presence of nitrite and clone their promoters as new sensors.
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    <li> be massively present on the sponge surface / avoiding being in an unfavorable position for food competition.</li>
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The following section describes protocols used to optimize (Rich media) & control (Minimal media) growth conditions.
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    <li> be found mainly in sponges / avoid spreading to species in contact with sponges.</li>
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    <li> be the phenotypically closest possible  to a known bacterium. / avoid cell cultures difficulties.</li>
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Latest revision as of 02:26, 18 October 2014

Pseudovibrio denitrificans the bacterium to modify



== From a poorly known genius, a jewel emerged ==



At first glance, working on the microbiome of sponges appears like a hazardous task: 5734 articles for marine sponges, only 51 with microbiome of sponges, only 28 articles mentioning Pseudovibrio genius, only 5 mentioning Pseudovibrio denitrificans, and finally none about a genetic engineering system in it.



Moreover with only 12 species in the genius, and 2 strains sequenced (Pseudovibrio sp. FO-BEG1, Pseudovibrio sp. JE062) we knew we would have to sequence our strain of Pseudovibrio denitrificans with a mapping to one of the 2 strains references. A formidable drawback even considering that in the first place the species was chosen because of its natural denitrification ability (see toxic compound) and easy to source at DMSZ.



On the other hand Pseudovibrio denitrificans, or strains related to the species, has been shown to be in the majority in at least six microbiomes of sponges in the Mediterranean Sea, where spongia officinalis resides.



Now the thorny question is: can it be easily used in our lab?

== A bacterium we could work with ==



In the literature Pseudovibrio denitrificans is described as a Gram-negative, motile by means of one to several lateral or subpolar flagella requiring NaCl for growth. It exhibits optimal growth at about 30 C°, pH 8 and 3 % NaCl, and a doubling time of 45 min in rich media. In a nutshell the conditions seem favourable for an iGEM project.



They are known to be capable of anaerobic growth by carrying out denitrifying metabolism using nitrate, nitrite or nitrous oxide as terminal electron acceptors. Being consequently a main reason for our RNAseq study to discover the transcripts upregulated by the presence of nitrite and clone their promoters as new sensors.



The following section describes protocols used to optimize (Rich media) & control (Minimal media) growth conditions.