Difference between revisions of "Team:Austin UTexas/Sandbox Page"

Line 3: Line 3:
 
<html>
 
<html>
 
<head>
 
<head>
−
<style>
+
<body>
−
body {
+
−
    background-color: #E0FFFF
+
  
−
}
+
<br>
  
−
</style>
+
<p style= "color:white; font-family: verdana">The indigenous gut microbiota in humans possesses the ability to synthesize neurotransmitters that are hypothesized to modulate behavioral, cognitive, and emotional processes of the body via the gut-brain axis (1). Since the emergence of this axis theory, researchers have been interested in genetically modifying probiotics for use as edible delivery vehicles of neurotransmitters to treat a variety of illnesses. There is limited experimental evidence regarding the effectiveness of these probiotics in the gut as this is a relatively novel concept. </p>
−
</head>
+
−
<div class="column full_size">
+
−
<div style="background-color: #282828;padding:14px;">
+
  
 
<br>
 
<br>
  
−
<div class="clear"></div>
+
<p style= "color:white; font-family: verdana">Thus, our project is aimed at engineering an effective probiotic capable of producing higher levels of gamma-aminobutyric acid (GABA) to treat patients with anxiety and certain bowel disorders. GABA is the neurotransmitter of interest due to its proposed anxiolytic, anti-hypertensive, and digestion-regulating effects (2-4). In order to create an accurate and translational model, we will utilize <i>Lactobacillus plantarum</i> as our probiotic, as it is native to the human gut (5). <i>L. plantarum</i> is also commonly found in a wide variety of foods, including yogurt, kimchi, and cheese, allowing us to easily utilize our genetically-modified version to create medicinal food (6-8). Although <i>L. plantarum</i> can naturally synthesize GABA, we intend to expand the GABA-producing potential of the bacteria by characterizing and subsequently optimizing the expression of glutamate decarboxylase, an enzyme responsible for converting glutamate into GABA (9). </p>
  
 +
<div class="column full_size_outer">
 +
<div class="column full_size_inner">
 +
 +
<style>
 +
body {
 +
  background: #282828;
 +
}
 +
</style>
 +
 +
<div class="column-left">
 +
<h4 style="font-size:22px;text-align: center; font-family: verdana">Part Plasmid Assembly</h4>
 
<br>
 
<br>
−
<div class="column full_size">
+
<a href="https://2017.igem.org/Team:Austin_UTexas/PartPlasmids"> <img src="https://static.igem.org/mediawiki/2017/4/4f/T--Austin_UTexas--gadBAndPromoterPartPlasmids.jpg" style="width:100%;"></a></center>
−
<h2 style= "color: lightcyan; text-align:center;font-family: verdana">Collaborations</h2>
+
<p>We had to construct various part plasmids in order to use them in Golden Gate Assembly.</p>
 
<br>
 
<br>
−
 
−
<p style="color: white; text-align:justify; font-family: verdana">To fulfill the silver medal requirement, the UT Austin team collaborated with the Texas Tech and Rice iGEM teams. The UT Austin team reached out to discuss ways in which all parties involved could cooperate and assist each other in order to enhance each team project.</p>
 
 
<br>
 
<br>
−
 
+
</div>
−
</html>
+
<div class="column-right">
−
https://static.igem.org/mediawiki/2017/2/2d/Igemmeetup.png
+
<h4 style="font-size:22px;text-align: center">Working with <i>L. plantarum</i></h4>
−
<html>
+
−
 
+
 
<br>
 
<br>
−
 
+
<br>
−
<p style="color: white; text-align:justify; font-family: verdana">In the spirit of collaboration, the UT Austin team reached out to both Rice and Texas Tech to formally invite both teams to participate in the University of Texas 2017 Fall Research Undergraduate Symposium. Upon this invitation, each team began electronically communicating with one another in order to brainstorm ways in which collaboration could benefit and improve each of the team projects.</p>
+
<a href="https://2017.igem.org/Team:Austin_UTexas/LPlantarum"> <img src="https://static.igem.org/mediawiki/2017/0/09/T--Austin_UTexas--LPlantarum.jpg" style="width:100%;"></a>
 +
<p>Our proposed bacteria for over expression of GABA in the human gut is <i>L. plantarum</i>, a probiotic..</p>
 +
</div>
 +
</div>
  
 
<br>
 
<br>
  
−
<p style="color: white; text-align:justify; font-family: verdana">Prior to the meetup, multiple ideas were brainstormed on determining how exactly each team could assist the other and vice-versa. On Saturday, September 23, 2017, iGEM teams from Texas Tech University, Rice University, and the University of Texas met at the Undergraduate Research Symposium in Austin, Texas. It was here that each team formally presented their projects to an audience and panel of judges as part of the program. After the symposium presentations concluded, all teams met to discuss specific collaboration details and provide feedback to one another. As a relatively new team, Texas Tech received extensive constructive criticism from team UT Austin, who overall has more experience in participating in the iGEM competition and completing the requirements. Texas Tech presented a draft of the initial PowerPoint presentation they were planning to showcase at the iGEM Jamboree. Darron Tharp, newcomer to the Texas Tech team and iGEM, was the main presenter of this presentation. First, Tharp presented their research slides in their entirety. After doing so, team UT Austin had Texas Tech go through every individual slide and gave them feedback on the content, formatting, and presentation technique utilized. In the process of this, Texas Tech asked various questions that UT Austin answered and clarified. This series of feedback with the presentation was repeated with team Rice. After both teams had their presentations assessed and critiqued by team UT Austin, the joint effort was continued by discussing a more direct means of collaborating.</p>
+
<h5 style="color:deepskyblue; font-family: verdana">References</h5>
  
−
<br>
+
<ol style="font-size:11px; font-family: verdana">
  
−
<p style="color: white; text-align:justify; font-family: verdana">Initially, there were talks of possibly having Texas Tech transform <i>Lactobacillus</i> with a different protocol to help troubleshoot issues our team was having in our original electroporation procedures.</p>
+
<li> Mayer E. A. et al. Gut Microbes and the Brain: Paradigm Shift in Neuroscience. <i>J Neurosci.</i>34(46): 15490-96 (2014). </li>
−
</div>
+
<li> Bravo, J. A. et al. Ingestion of <i>Lactobacillus</i> strain regulates emotional behavior and central GABA receptor expression in a mouse via the vagus nerve. <i>Proc Natl Acad Sci USA</i>. 108(38): 16050-5 (2011).</li>
 +
<li> Shimada M. et al Anti-hypertensive effect of gamma-aminobutyric acid (GABA)- rich <i>Chlorella</i> on high-normal blood pressure and borderline hypertension in placebo-controlled double blind study. <i>Clin Exp Hypertens</i>. 21(4): 342-54 (2009).</li>
 +
<li> Auteri M. et al. GABA and GABA receptors in the gastrointestinal tract: from motility to inflammation. <i>Pharmacological Research</i>. 93: 11-21 (2015).</li>
 +
<li> De Vries, M. et al. <i>Lactobacillus plantarum</i> - survival, functional and potential probiotic properties in the human intestinal tract. <i>International Dairy Journal</i>. 16(0): 1018-1028 (2006).</li>
 +
<li> Shan, Y. et al. Evaluation of improved gamma-aminobutyric acid production in yogurt using <i>Lactobacillus plantarum</i> NDC75017. <i>J Dairy Sci</i>. 98(4): 2138-49 (2015).</li>
 +
<li> Siragusa, S. et al. Synthesis of gamma-aminobutyric acid by lactic acid bacteria isolated from a variety of italian cheeses. <i>Appl Environ Microbiol</i>. 73(22): 7283-90 (2007).</li>
 +
<li> Yoon, J. H. <i>Lactobacillus kimchii</i> sp. Nov., a new species from kimchi. <i>Int J Syst Evol Microbiol</i>. 50: 1789-95 (2000).</li>
 +
<li> Yunes, R. A. GABA production and structure of <i>gadB</i>/<i>gadC</i> genes in <i>Lactobacillus</i> and <i>Bifidobacterium</i> strains from human microbiota. <i>Anaerobe</i>. 42: 197-204 (2016).</li>
 +
 
 +
</ol>
  
 +
</body> 
  
 
</html>
 
</html>

Revision as of 15:22, 30 October 2017


The indigenous gut microbiota in humans possesses the ability to synthesize neurotransmitters that are hypothesized to modulate behavioral, cognitive, and emotional processes of the body via the gut-brain axis (1). Since the emergence of this axis theory, researchers have been interested in genetically modifying probiotics for use as edible delivery vehicles of neurotransmitters to treat a variety of illnesses. There is limited experimental evidence regarding the effectiveness of these probiotics in the gut as this is a relatively novel concept.


Thus, our project is aimed at engineering an effective probiotic capable of producing higher levels of gamma-aminobutyric acid (GABA) to treat patients with anxiety and certain bowel disorders. GABA is the neurotransmitter of interest due to its proposed anxiolytic, anti-hypertensive, and digestion-regulating effects (2-4). In order to create an accurate and translational model, we will utilize Lactobacillus plantarum as our probiotic, as it is native to the human gut (5). L. plantarum is also commonly found in a wide variety of foods, including yogurt, kimchi, and cheese, allowing us to easily utilize our genetically-modified version to create medicinal food (6-8). Although L. plantarum can naturally synthesize GABA, we intend to expand the GABA-producing potential of the bacteria by characterizing and subsequently optimizing the expression of glutamate decarboxylase, an enzyme responsible for converting glutamate into GABA (9).

Part Plasmid Assembly


We had to construct various part plasmids in order to use them in Golden Gate Assembly.



Working with L. plantarum



Our proposed bacteria for over expression of GABA in the human gut is L. plantarum, a probiotic..


References
  1. Mayer E. A. et al. Gut Microbes and the Brain: Paradigm Shift in Neuroscience. J Neurosci.34(46): 15490-96 (2014).
  2. Bravo, J. A. et al. Ingestion of Lactobacillus strain regulates emotional behavior and central GABA receptor expression in a mouse via the vagus nerve. Proc Natl Acad Sci USA. 108(38): 16050-5 (2011).
  3. Shimada M. et al Anti-hypertensive effect of gamma-aminobutyric acid (GABA)- rich Chlorella on high-normal blood pressure and borderline hypertension in placebo-controlled double blind study. Clin Exp Hypertens. 21(4): 342-54 (2009).
  4. Auteri M. et al. GABA and GABA receptors in the gastrointestinal tract: from motility to inflammation. Pharmacological Research. 93: 11-21 (2015).
  5. De Vries, M. et al. Lactobacillus plantarum - survival, functional and potential probiotic properties in the human intestinal tract. International Dairy Journal. 16(0): 1018-1028 (2006).
  6. Shan, Y. et al. Evaluation of improved gamma-aminobutyric acid production in yogurt using Lactobacillus plantarum NDC75017. J Dairy Sci. 98(4): 2138-49 (2015).
  7. Siragusa, S. et al. Synthesis of gamma-aminobutyric acid by lactic acid bacteria isolated from a variety of italian cheeses. Appl Environ Microbiol. 73(22): 7283-90 (2007).
  8. Yoon, J. H. Lactobacillus kimchii sp. Nov., a new species from kimchi. Int J Syst Evol Microbiol. 50: 1789-95 (2000).
  9. Yunes, R. A. GABA production and structure of gadB/gadC genes in Lactobacillus and Bifidobacterium strains from human microbiota. Anaerobe. 42: 197-204 (2016).