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What is the context of this research?

Using the native sludge bacteria Nitrosospira multiformis and Nitrosomonas eutropha as chassis we inserted genes to produce erythromycin esterase B and β-lactamase to breakdown azythromycin and penicillin. We also inserted nirS, norB, norC, and nosZ from Pseudomonas aeruginosa PAO1 to convert nitrates into nitrogen gas, as well as genes to produce dispersin, amylase, and AHL-lactonase to inhibit the biofilm formation which blocks helpful bacteria from functioning fully. To increase bacteriophage resistance, prophage in the Nitrosospira and Nitrosomonas genomes were identified and used to build a guide RNA region for a Type II CRISPR system. These improvements will help reduce antibiotic resistance, increase water reclamation, prevent algal blooms, and allow more biomass to be harvested.

What is the significance of this project?

Wastewater facilities face challenges in effectively processing waste including residual antibiotics, excess nitrates, biofilm buildup and low survival rates of microbes essential to biodegradation. Our work will provide more effective solutions to handling these issues.

What are the goals of the project?

1. Improve bacteriophage resistance 1000-fold in N. multiformis and characterize the CRISPR system allowing for this increased resistance. This will be the first time this particular CRISPR is being characterized.
2. Introduce denitrification genes into N. multiformis to convert ammonia into nitrogen gas. This will allows us to remove excess nitrates from sewage effluent and prevent eutrophication.
3. Introduce genes into N. multiformis to degrade two of the most common classes of antibiotics found in wastewater.
4. Test efficacy of genes that degrade biofilms that we have introduced into N. multiformis.
5. Test genes that will eliminate our modified bacteria if they leave the wastewater (acts as a control).
6. Present our research at the international iGem jamboree!

Requirements

Please be sure to keep these links, your audience will want to find your:

• Home
• Team
• Official Team Profile
• Project
• Parts
• Modeling
• Notebook
• Safety
• Attributions

There are a few wiki requirements teams must follow:

• All pages, images and files must be hosted on the 2014.igem.org server.
• All pages must be created under the team’s name space.
• As part of your documentation, keep the links from the menu to the left.
• Do not use flash in wiki code.
• The iGEM logo should be placed on the upper part of every page and should link to 2014.igem.org.

Visit the Wiki How To page for a complete list of requirements, tips and other useful information.

Tips

We are currently working on providing teams with some easy to use design templates.
In the meantime you can also view other team wikis for inspiration! Here are some very good examples

• 2013 SDU Denmark
• 2013 SYSU China
• 2013 Shenxhen BGIG ATCG
• 2013 Colombia Unianades
• 2013 Lethbridge

For a full wiki list, you can visit iGEM 2013 web sites and iGEM 2012 web sites lists.

This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started:

• State your accomplishments! Tell people what you have achieved from the start.
• Be clear about what you are doing and what you plan to do.
• You have a global audience! Consider the different backgrounds that your users come from.
• Make sure information is easy to find; nothing should be more than 3 clicks away.
• Avoid using very small fonts and low contrast colors; information should be easy to read.
• Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the iGEM 2013 calendar
• Have lots of fun!