Rsdbenitez (Talk | contribs) |
Rsdbenitez (Talk | contribs) |
||
(9 intermediate revisions by the same user not shown) | |||
Line 827: | Line 827: | ||
letter-spacing: 0.5px; | letter-spacing: 0.5px; | ||
text-align: center; | text-align: center; | ||
− | + | margin: 1.5em 0 0 0; | |
} | } | ||
h4 { | h4 { | ||
Line 2,882: | Line 2,882: | ||
<div class="expandable-content-wrap expandable-content-wrap-3"> | <div class="expandable-content-wrap expandable-content-wrap-3"> | ||
<div class="expandable-content"> | <div class="expandable-content"> | ||
− | <h3> | + | <h3>Forming Our Team</h3> |
− | <p> | + | <p>Prokhor, Stephen and Rachel are new members joining our team. We've all come together for our first meeting to discuss potential project ideas and go over the general timeline for the year. Everyone has their own specific interests ranging from plant physiology to wine microbiology - it's great to see a diverse range of options. We've identified waterway pollution as a huge problem in New Zealand: each year we're seeing more incidences of toxic algal bloom formation in the summer and have begun searching potential ways to use synthetic biology to combat this issue. We're also interested in the idea of creating a 'resurrection plant', capable of tolerating desiccation using trehalose-induced autophagosomal activity.</p> |
<img src="https://static.igem.org/mediawiki/2018/f/f3/T--Auckland_MOD--lab.jpg"> | <img src="https://static.igem.org/mediawiki/2018/f/f3/T--Auckland_MOD--lab.jpg"> | ||
− | + | <p></p> | |
− | <p> | + | |
</div> | </div> | ||
</div> | </div> | ||
Line 2,902: | Line 2,901: | ||
<div class="expandable-content"> | <div class="expandable-content"> | ||
<h3>Refining our Biological Solution</h3> | <h3>Refining our Biological Solution</h3> | ||
− | <p>After carrying out our own research, we've looked into the effects of dairy run-off entering the water system. Discussion with our PI and researchers within the university has helped narrow down the chosen metabolic pathway we might upregulate | + | <p>We've decided as a team to advance our nitrogen project idea. After carrying out our own research, we've looked into the effects of dairy run-off entering the water system and where our plant might intercept this cycle. Discussion with our PI and researchers within the university has helped narrow down the chosen metabolic pathway we might upregulate - we've chosen <i>Arabidopsis thaliana</i> as our model organism and plan on increasing the uptake and breakdown of excess urea directly in the soil.</p> |
− | <p>Set up a meeting with Dr Nijat Imin who specialises in nitrogen activity in plants. We were already thinking of upregulating <b>AtDUR3</b> urea transporter and incorporating H. pylori’s | + | <p>Set up a meeting with Dr Nijat Imin who specialises in nitrogen activity in plants. We were already thinking of upregulating <b>AtDUR3</b> urea transporter and incorporating H. pylori’s high affinity <b>urease</b>. He's informed us that the produced ammonia is toxic in high concentrations and suggested a few transporters that would be able to redirect it to other parts of the plant. He is excited about the project and is willing to give us more support throughout the year.<p> |
<img src="https://static.igem.org/mediawiki/2018/f/f3/T--Auckland_MOD--lab.jpg"> | <img src="https://static.igem.org/mediawiki/2018/f/f3/T--Auckland_MOD--lab.jpg"> | ||
− | <p> <b>Lab induction time!</b> We've all had to complete our safety training so we can jump into the PC1 facilities for our lab work. </p> | + | <p> <b>Lab induction time!</b> We've all had to complete our safety training, led by our PI Austen Ganley, so we can jump into the PC1 facilities for our lab work. </p> |
<h4>Looking Beyond the Lab</h4> | <h4>Looking Beyond the Lab</h4> | ||
− | <p>If we're looking at a product for dairy farmers, it's important to start thinking about market validation. Rachel has begun speaking to social scientists in the university to get an idea of how to conduct a focus group, and how to engage with students as part of our community engagement efforts.</p> | + | <p>If we're looking at a product for dairy farmers, it's important to start thinking about market validation. Stephen has been looking into existing run off management practices. Rachel has begun speaking to social scientists in the university to get an idea of how to conduct a focus group, and how to engage with students as part of our community engagement efforts. </p> |
</div> | </div> | ||
</div> | </div> | ||
Line 2,924: | Line 2,923: | ||
<h3>Identifying the Urea Metabolic Pathway</h3> | <h3>Identifying the Urea Metabolic Pathway</h3> | ||
<p>The team met with Nijat again after researching further into his suggestions. <i>H. pylori</i> high affinity urease will be a bit difficult to introduce as it would need special approval to be inserted into <i>A. thaliana</i>. It's also a human pathogen and has a different accessory proteins (so it's very complicated!). We need to look into another way of upregulating urease. </p> | <p>The team met with Nijat again after researching further into his suggestions. <i>H. pylori</i> high affinity urease will be a bit difficult to introduce as it would need special approval to be inserted into <i>A. thaliana</i>. It's also a human pathogen and has a different accessory proteins (so it's very complicated!). We need to look into another way of upregulating urease. </p> | ||
− | <p> We’ve decided that we want to upregulate the <b>AtDUR3</b> gene then the urease or rate-limiting accessory protein. There is limited research around upregulating urease activity: one of the urease accessory proteins, <b>UreG</b>, might be our best luck as an alternative to expressing urease with all accessory genes. | + | <p> We’ve decided that we want to upregulate the <b>AtDUR3</b> gene then the urease or rate-limiting accessory protein. There is limited research around upregulating urease activity: one of the urease accessory proteins, <b>UreG</b>, might be our best luck as an alternative to expressing urease with all accessory genes. Rachel and Stephen have been researching which glutamine synthetase gene we might upregulate. <b>GS1;2 isogene</b> appears to be the most successful in producing glutamate from ammonia at the highest capacity. </p> |
− | + | <h3> Beginning Work in the Plant Lab</h3> | |
− | <h3> Beginning Work in the Plant Lab<h3> | + | <p> We've selected Greta, Prokhor and Stephen to undergo the plant lab induction in mid-May. They will be responsible for growing our <i>A. thaliana</i> plants in this PC2 lab, with aid of the lab supervisor, Nathan Deed. </p> |
− | + | ||
<p><b>Late May:</b> We received training for entry procedures, plant sowing, logging our sowed/bleached plants, and seed sterilisation. We've successfully sowed our first round of 24 wild-type <i> A. thaliana</i> seeds. We watered the plants with 1:10 hydroponics solution and recorded planting dates. </p> | <p><b>Late May:</b> We received training for entry procedures, plant sowing, logging our sowed/bleached plants, and seed sterilisation. We've successfully sowed our first round of 24 wild-type <i> A. thaliana</i> seeds. We watered the plants with 1:10 hydroponics solution and recorded planting dates. </p> | ||
<img src="https://static.igem.org/mediawiki/2018/9/9f/T--Auckland_MOD--StJohns_ProkhorSneha1.jpeg"> | <img src="https://static.igem.org/mediawiki/2018/9/9f/T--Auckland_MOD--StJohns_ProkhorSneha1.jpeg"> | ||
Line 2,953: | Line 2,951: | ||
<p> <b> Early June: </b> Our first algal bloom has been reported. As a result we've cleaned and exchanged our trays for new clean ones. We've been continuously watering our plants and recording growth every few days. </p> | <p> <b> Early June: </b> Our first algal bloom has been reported. As a result we've cleaned and exchanged our trays for new clean ones. We've been continuously watering our plants and recording growth every few days. </p> | ||
<p> <b> Rest of June: </b> We've had reports of yellowing leaves, indicating stress to the plants. We've been advised to apply hydroponic solution directly to the yellowing plants. The plant facilities had a heating malfunction on the 25th, so we had to move them to the temperature controlled room. No plant damage was observed. </p> | <p> <b> Rest of June: </b> We've had reports of yellowing leaves, indicating stress to the plants. We've been advised to apply hydroponic solution directly to the yellowing plants. The plant facilities had a heating malfunction on the 25th, so we had to move them to the temperature controlled room. No plant damage was observed. </p> | ||
+ | <p> | ||
<h4>Getting started with our BioBrick Characterisation</h4> | <h4>Getting started with our BioBrick Characterisation</h4> | ||
− | <p> <b> Early June: </b> Had a meeting with 2017 iGEM team lab members, Judith and Jess, to get a better idea of how we can work more efficiently. They suggested not to continue from their BioBrick project this year, but instead do a fluorescent or chromoprotein. We've chosen <b>BBa_E0020</b> from 2004 - we’ll be finding out at what pH we observe the highest fluorescent intensity. | + | <p> <b> Early June: </b> Had a meeting with 2017 iGEM team lab members, Judith and Jess, to get a better idea of how we can work more efficiently. They suggested not to continue from their BioBrick project this year, but instead do a fluorescent or chromoprotein. We've chosen <b>BBa_E0020</b> from 2004 - we’ll be finding out at what pH we observe the highest fluorescent intensity. We've met with our lab supervisor, Jasper, to go over our lab protocol and familiarise ourselves to the BioBrick standard system. </p> |
− | + | ||
<img src="https://static.igem.org/mediawiki/2018/7/73/T--Auckland_MOD--PC1_StackedPlates.jpeg"> | <img src="https://static.igem.org/mediawiki/2018/7/73/T--Auckland_MOD--PC1_StackedPlates.jpeg"> | ||
<p><b> Mid-June: </b> Finally jumping into the lab to do our plating and preparatory steps for the transformation into competent <i>E. coli</i> strain, DH5α. We’re all starting to feel the pressure of upcoming exams which is getting in the way of lab progress. Right now, the priority is to grow our transformed <i>E. coli</i> and get them on ice so we can experiment on them later in July. </p> | <p><b> Mid-June: </b> Finally jumping into the lab to do our plating and preparatory steps for the transformation into competent <i>E. coli</i> strain, DH5α. We’re all starting to feel the pressure of upcoming exams which is getting in the way of lab progress. Right now, the priority is to grow our transformed <i>E. coli</i> and get them on ice so we can experiment on them later in July. </p> | ||
− | |||
<img src="https://static.igem.org/mediawiki/2018/9/95/T--Auckland_MOD--CFP_PCRgel1.jpeg"> | <img src="https://static.igem.org/mediawiki/2018/9/95/T--Auckland_MOD--CFP_PCRgel1.jpeg"> | ||
+ | <p> Our first transformation of our BioBrick has been successful! We’re now waiting for our primers (from our IDT free DNA supply) to arrive in the mail so we can run colony PCR and confirm our insert is being expressed. </p> | ||
</div> | </div> | ||
</div> | </div> | ||
Line 2,979: | Line 2,977: | ||
<h4> Progress in the Plant Lab</h4> | <h4> Progress in the Plant Lab</h4> | ||
<p> In early July, Greta and Prokhor and completed the training for staking and bagging of our Arabidopsis plants with sprouted stems. This is done to contain the seed pods and prevent unintentional spreading of GMOs. We've also been trimming lateral shoots to optimize growth. Continuous management of algal formation and applying hydroponic solution in response to plant growth. On the 27th, the plant lab team had their seed collection training, involving the cutting of dried plant stems, freeing the seeds from their pods and collecting them for storage.</p> | <p> In early July, Greta and Prokhor and completed the training for staking and bagging of our Arabidopsis plants with sprouted stems. This is done to contain the seed pods and prevent unintentional spreading of GMOs. We've also been trimming lateral shoots to optimize growth. Continuous management of algal formation and applying hydroponic solution in response to plant growth. On the 27th, the plant lab team had their seed collection training, involving the cutting of dried plant stems, freeing the seeds from their pods and collecting them for storage.</p> | ||
− | <p> The UreG plasmid the plant lab ordered from another lab group arrived in the mail. We want to test our Gateway protocol as we wait for our IDT vectors to arrive, but first we need to find out how to add the <i>attB</i> sites to our plasmid. </p> | + | <p> The UreG plasmid the plant lab ordered from another lab group arrived in the mail. We want to test our Gateway protocol as we wait for our IDT vectors to arrive, but first we need to find out how to add the <i>attB</i> sites to our plasmid. We've also been contacting the Witte research group to get a sample of their DUR3 transgenic seeds however this seems unlikely. We will instead receive their plasmid - this set back means we only have time to insert two gene constructs into our plant, which may not be enough to increase metabolic flux of urea uptake. We'll still carry on upregulating the DUR3 transporter and the glutamine synthetase enzyme in the pathway and see what happens. We've designed the GS1;2 construct with the help of Nijat and Matthew, and ordered it through IDT.</p> |
<img src="https://static.igem.org/mediawiki/2018/a/a7/T--Auckland_MOD--SnehaPlating.jpeg"> | <img src="https://static.igem.org/mediawiki/2018/a/a7/T--Auckland_MOD--SnehaPlating.jpeg"> | ||
<h3>Working through the BioBrick lab</h3> | <h3>Working through the BioBrick lab</h3> | ||
Line 2,992: | Line 2,990: | ||
<h4> Expanding the iGEM Community</h4> | <h4> Expanding the iGEM Community</h4> | ||
<p> One of our contribution initiatives is to establish another iGEM team in New Zealand. We've spoken to Kyle Webster, a PhD student previously from Canterbury University, who was involved in our past iGEM teams here in Auckland. He's keen to help us get in contact with the right people in Canterbury University to spread this opportunity to other undergrads down in the South Island.</p> | <p> One of our contribution initiatives is to establish another iGEM team in New Zealand. We've spoken to Kyle Webster, a PhD student previously from Canterbury University, who was involved in our past iGEM teams here in Auckland. He's keen to help us get in contact with the right people in Canterbury University to spread this opportunity to other undergrads down in the South Island.</p> | ||
+ | <p> Sneha, Greta and Rachel also went to Papakura High School to meet their STEM teacher and begin planning a soil microbiology session with the students. We want to add onto their current hydroponics project by looking at the microbial diversity in their system and the roles they play. We also had a good discussion about empowering students from low decile schools about science enquiry and sustainability practices which they can apply to their own experiences. </p> | ||
</div> | </div> | ||
</div> | </div> | ||
Line 3,006: | Line 3,005: | ||
<div class="expandable-content-wrap expandable-content-wrap-8"> | <div class="expandable-content-wrap expandable-content-wrap-8"> | ||
<div class="expandable-content"> | <div class="expandable-content"> | ||
− | <h3> | + | <h3>Progressing in the Plant Lab</h3> |
− | <p> | + | <p>Our synthesized GS1 construct as arrived from IDT and we've begun the Gateway reactions to get this into our final pB2GW7 vector. So far we have only reached the BP reaction - we're having trouble optimizing our PCR protocol and enzyme digest to check the reaction has worked. With help from our supervisor Matthew, we've been troubleshooting our failed experiments. All part of the learning experience!</p> |
− | <img src="https://static.igem.org/mediawiki/2018/ | + | <img src="https://static.igem.org/mediawiki/2018/6/61/T--Auckland_MOD--ProkhorLaminarFlow.jpeg"> |
− | <h4> | + | <h4>Preparing for the Jamboree</h4> |
− | <p> | + | <p>Our main pressure point is sourcing the funding from our investors to get tickets to Boston and attend the Jamboree in October. We've been contacting biotech firms in the Auckland region and research institutes around New Zealand who have similar interests to our project, whether they include plant biology or science commercialisation.</p> |
</div> | </div> | ||
</div> | </div> |
Latest revision as of 02:35, 5 September 2018
j<!DOCTYPE html>
Notebook
It's been a busy year! Take a look at the progress we've been making in our project: both in and outside of the lab.
"Competing in iGEM required more strength, endurance, and sacrifice than any other project I have undertaken. But nothing could have prepared me more for the post graduate study I am now pursuing than formulating and completing our project as an independent scienctist."
Judith Glasson, Alumni
Hold to view image
Keen to talk?
If you're interested, have questions, or want to know more, don't hesitate to contact us directly.