Difference between revisions of "Team:Montpellier/L jensenii"

 
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<img class="banniere" src="https://static.igem.org/mediawiki/2018/a/ac/T--Montpellier--banniere_lactobacillus_jensenii.png"/>
  
 
<section>
 
<section>
  
<h1>How to work with <i>Lactobacillus jensenii</i>?</h1>
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<p>For our project, we decided to study <i>Lactobacillus jensenii</i> (Figure 1), as it appeared to be a promising candidate for several reasons.</p>
 
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<p>This toolbox contains all kind of information that can be useful for working with <i>Lactobacillus jensenii</i>. First, we provide culture conditions that we used and were proven to be working. Second, it contains a transformation protocol that we optimized and found to be the most efficient of all those we tested. Finally, the toolbox contains all DNA sequences (vectors and biobricks parts including <i>L. jensenii</i> specific promoters) that we used in <i>L. jensenii</i> for our project. We hope that this toolbox will facilitate the use of <i>L. jensenii</i> by other iGEM and research teams. We plan to extend the toolbox in the future by providing the community with a physical distribution of well-characterized vectors and regulatory elements.</p>
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<h2>How to grow <i>Lactobacillus jensenii</i>?</h2><hr/>
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<p>For our project we used the following strain and culture medium :</p>
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<h4>Bacteria</h4>
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<p><a class="lien" href="http://www.lgcstandards-atcc.org/products/all/25258.aspx?geo_country=fr" target="_blank"><i>Lactobacillus jensenii</i> Gasser et al. (ATCC® 25258™)</a></p>
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<h4>Culture</h4>
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<p><a class="lien" href="https://www.fishersci.fr/shop/products/bd-difco-dehydrated-culture-media-lactobacilli-mrs-broth-3/11713553#?keyword=BD+288130&change_lang=true" target="_blank">BD Difco™ <i>Lactobacilli</i> MRS Broth</a></p>
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<p>We used this broth for both our liquid and solid culture media. The liquid medium was used to do the solid medium by adding agar to it.  We tested different concentrations of Agar to measure which one was the best.</p>
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<img class="img_plate" src="https://static.igem.org/mediawiki/2018/9/97/T--Montpellier--agar5_mtp.jpg"/>
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<img class="image_figure_50" src="https://static.igem.org/mediawiki/2018/3/34/T--Montpellier--lactobacillus_jensenii.jpg"/><br/>
<p>5g/L</p>
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<figcaption><span class="underline">Figure 1:</span> <i>L. jensenii</i> strain used for our project under 100x magnification.</figcaption>
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<div class="image_plate">
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<figure class="floating">
<img class="img_plate" src="https://static.igem.org/mediawiki/2018/8/87/T--Montpellier--agar7_mtp.jpg"/>
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<img class="image_figure_70" src="https://static.igem.org/mediawiki/2018/7/7d/T--Montpellier--vagina_compostion2_mtp.png"/><!--float left-->
<p>7g/L</p>
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<figcaption><span class="underline">Figure 2:</span> Representation of the vaginal flora composition for the majority of women.</figcaption>
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<div class="image_plate">
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<p><i>L. jensenii</i> is a normal inhabitant of the lower reproductive tract in healthy women. More than 20 species have been detected in the vagina, but in healthy conditions only one or two of the four main species (mainly <i>L. crispatus</i> and <i>L. iners</i>, but also <i>L. jensenii</i> and <i>L. gasseri</i>) are dominant (Figure 2) <a class="lien" href="#references">[1]</a><a class="lien" href="#references">[2]</a>.</p>
<img class="img_plate" src="https://static.igem.org/mediawiki/2018/3/30/T--Montpellier--agar10_mtp.jpg"/>
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<p>10g/L</p>
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</div>
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<div class="image_plate">
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<p>While gynecological studies have found that bacterial population makeup differs around the world (e.g. <i>L. crispatus</i> and <i>L. jensenii</i> are dominant in Caucasian populations while <i>L. crispatus</i> and <i>L. gasseri</i> are dominant in women of Japanese descent) <a class="lien" href="#references">[3]</a>, we believe that the work we do towards developing <i>L. jensenii</i> as an engineering platform will be transferable to similar species. Indeed, if we can demonstrate our concept using <i>L. jensenii</i>,  the same methodology will likely be applicable to other inhabitants of the vaginal microbiota in an effort to improve the effects of our non-hormonal contraception system.</p>
<img class="img_plate" src="https://static.igem.org/mediawiki/2018/7/7e/T--Montpellier--agar12_mtp.jpg"/>
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<p>12g/L</p>
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</div>
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<div class="image_plate">
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<p>Discovered by F. Gasser, M. Mandel, and M. Rogosa in 1969 <a class="lien" href="#references">[4]</a>, <i>L. jensenii</i> is currently used very infrequently in synthetic biology. This bacterium has not been well characterized and few research teams have been working with this strain in the last few decades. Finding information for how to work with this bacterium was arduous for the team. However, we are convinced that future work on this strain can be crucial for women’s health. This is why we decided to create a toolbox for working with <i>Lactobacillus jensenii</i>.
<img class="img_plate" src="https://static.igem.org/mediawiki/2018/a/a4/T--Montpellier--agar15_mtp.jpg"/>
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<p>15g/L</p>
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</div>
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<figcaption><span class="underline">Figure 1:</span> Plates with <i>L. jensenii</i> grown for 24 hours with different concentrations of Agar.</figcation>
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</section>
  
</div>
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<section class="references" id="references">
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  <table class="references_table">
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    <tr>
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      <th class="references_title" colspan="2">References</th>
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    </tr>
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<tr>
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      <td class="references_left">[1]</td>
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      <td class="references_right">Reid, G., McGroarty, J. A., Tomeczek, L., & Bruce, A. W. (1996). Identification and plasmid profiles of <i>Lactobacillus</i> species from the vagina of 100 healthy women. <i>FEMS Immunology & Medical Microbiology</i>, 15(1), 23-26.</td>
 +
    </tr>
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    <tr>
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      <td class="references_left">[2]</td>
 +
      <td class="references_right">Zhou, X., Bent, S. J., Schneider, M. G., Davis, C. C., Islam, M. R., & Forney, L. J. (2004). Characterization of vaginal microbial communities in adult healthy women using cultivation-independent methods. <i>Microbiology</i>, 150(8), 2565-2573.</td>
 +
    </tr>
 +
    <tr>
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      <td class="references_left">[3]</td>
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      <td class="references_right">Ravel, J., Gajer, P., Abdo, Z., Schneider, G. M., Koenig, S. S., McCulle, S. L., ... & Brotman, R. M. (2011). Vaginal microbiome of reproductive-age women. <i>Proceedings of the National Academy of Sciences, 108</i>(Supplement 1), 4680-4687.</td>
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    </tr>
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    <tr>
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      <td class="references_bottom_left">[4]</td>
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      <td class="references_bottom_right">Gasser, F., Mandel, M., & Rogosa, M. (1970). <i>Lactobacillus jensenii</i> sp. nov., a new representative of the subgenus Thermobacterium. <i>Microbiology</i>, 62(2), 219-222.</td>
 +
    </tr>
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  </table>
  
 
</section>
 
</section>

Latest revision as of 21:31, 16 October 2018

For our project, we decided to study Lactobacillus jensenii (Figure 1), as it appeared to be a promising candidate for several reasons.


Figure 1: L. jensenii strain used for our project under 100x magnification.
Figure 2: Representation of the vaginal flora composition for the majority of women.

L. jensenii is a normal inhabitant of the lower reproductive tract in healthy women. More than 20 species have been detected in the vagina, but in healthy conditions only one or two of the four main species (mainly L. crispatus and L. iners, but also L. jensenii and L. gasseri) are dominant (Figure 2) [1][2].

While gynecological studies have found that bacterial population makeup differs around the world (e.g. L. crispatus and L. jensenii are dominant in Caucasian populations while L. crispatus and L. gasseri are dominant in women of Japanese descent) [3], we believe that the work we do towards developing L. jensenii as an engineering platform will be transferable to similar species. Indeed, if we can demonstrate our concept using L. jensenii, the same methodology will likely be applicable to other inhabitants of the vaginal microbiota in an effort to improve the effects of our non-hormonal contraception system.

Discovered by F. Gasser, M. Mandel, and M. Rogosa in 1969 [4], L. jensenii is currently used very infrequently in synthetic biology. This bacterium has not been well characterized and few research teams have been working with this strain in the last few decades. Finding information for how to work with this bacterium was arduous for the team. However, we are convinced that future work on this strain can be crucial for women’s health. This is why we decided to create a toolbox for working with Lactobacillus jensenii.

References
[1] Reid, G., McGroarty, J. A., Tomeczek, L., & Bruce, A. W. (1996). Identification and plasmid profiles of Lactobacillus species from the vagina of 100 healthy women. FEMS Immunology & Medical Microbiology, 15(1), 23-26.
[2] Zhou, X., Bent, S. J., Schneider, M. G., Davis, C. C., Islam, M. R., & Forney, L. J. (2004). Characterization of vaginal microbial communities in adult healthy women using cultivation-independent methods. Microbiology, 150(8), 2565-2573.
[3] Ravel, J., Gajer, P., Abdo, Z., Schneider, G. M., Koenig, S. S., McCulle, S. L., ... & Brotman, R. M. (2011). Vaginal microbiome of reproductive-age women. Proceedings of the National Academy of Sciences, 108(Supplement 1), 4680-4687.
[4] Gasser, F., Mandel, M., & Rogosa, M. (1970). Lactobacillus jensenii sp. nov., a new representative of the subgenus Thermobacterium. Microbiology, 62(2), 219-222.