Team:Marburg/test

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Establishing Vibrio natriegens as the new chassis organism for synthetic biology.
With a doubling time of 7 minutes, we were able to complete a full cloning cycle in less than 12 hours!

Strain Engineering

We established three new strains derived from the original wildtype strain as chassis organisms for cloning, protein expression and protein interaction studies. Moreover, we got first evidence for a CRISPR/Cas9 mediated knockout.

Marburg Collection

We created the Marburg Collection, a highly flexible golden-gate based cloning toolbox consisting of 123 individual parts. Our novel measurement workflow was applied to obtain highly reproducible data on the behavior of our parts in V. natriegens.

Metabolic Engineering

We developed a workflow for accelerating metabolic engineering. As a proof of concept we established the first synthetic pathway in Vibrio natriegens to produce the platform chemical 3-hydroxypropionic acid.

Interlab

To collaborate with other teams and spread the word to other scientists on Vibrio natriegens' advantages we had teams from all over Europe conduct a growth curve or a InterLab inspired experiment.

Accessible Science

This Wiki is designed to be accessible for everyone! For this years Human Practices, iGEM Marburg decided to partner up with the BLISTA again. The BLISTA is the only Highschool for visually impaired students in Europe. As part of our project we (and other teams) designed a barrier-free website to take this step to remove barriers in communication!

Medal Criteria

Bronze

We fulfilled all bronze medal criteria. Our team registered for iGEM, we had a challenging but great iGEM season and we are really looking forward to attend the Giant Jamboree.

Deliverables

We already finalized our Wiki and Judging Form. We will hold our presentation at the Giant Jamboree in Boston and we expect exciting discussions at our poster.

Attributions

Our project was only possible with a great team and we also want to thank people, institutions and companies who helped with financial and infrastructural support or scientific advice, and we are grateful to honor them on our attributions page.

Characterization

We participated in the fifth iGEM InterLab measurement study, submitted our data in time and we are happy that our results were accepted by the measurement committee.

Silver

We are certain that we completed all silver medal criteria

Validated Part

We submitted BBa_K2560069 as a new basic parts. This part fulfills all formal requirements and was submitted to the registry. BBa_K2560069 is a 5’ Connector which was newly designed by us. We validated its function by showing that it can reduce crosstalk from backbone features 14 fold.

Collaborations

We strengthened the iGEM community in Germany by hosting the German iGEM Meetup and we planned and realized the Vibrigens InterLab study with eleven participating teams.

Human Practices

We got into contact with the BLISTA as part of our local community to open our project for everyone and we achieved to present our project in an accessible manner for the visually impaired.

Gold

We hope that we can convince you that we fulfilled all gold medal criteria.

Integrated Human Practices

We integrated valuable input from society, academia and industry into the design and execution of our project by starting a wiki collaboration and implementing the suggestions of stakeholders into the design of our strains.

Improve a previous part

We improved BBa_P10500 by replacing the dropout with a sfGFP cassette, resulting in BBa_K2560002.

Model Your Project

Using modeling we predicted the metabolic pathway that yields optimal 3HPA producting and conducted foundational work towards a novel pathway.

Demonstration of Your Work

We succeeded in all major parts of our project. We established genome engineering methods, created a toolbox and showed production of 3-HPA.

Special Prizes

We are proud about what we achieved and applied for the following special awards:

Integrated Human Practices

Education and Public Engagement

Model

Measurement

Software Tool

Measurement

Best New Basic Part

Best New Composite Part

Best Part Collection

Results	Achieved
Initially, we carried out foundational experiments to characterize the growth rate, pH, salt and antibiotic tolerance of V. natriegens and sequenced its genome.
We performed Cloning in One day: From transformation to isolated plasmids in less than 12 hours!
Low amounts of DNA could be transformed reliably and we successfully demonstrated challenging clonings such as Gibson Assembly with 5 and Aquacloning with 3 fragments as well as 8 part golden-gate reactions.
We designed and constructed the Marburg Collection, a highly flexible golden-gate based toolbox consisting of 123 parts.
Dozens of test constructs were built and tested to obtain characterization data for all part categories in our toolbox.
This was only possible with our novel experimental and data analysis workflow using platereader measurements.
We successfully demonstrated a genome engineering workflow in V. natriegens to establish three new lab strains for diverse applications.
Subsequently, we were able to use Flp/FRT recombinase system for marker recycling thus allowing additional rounds of genomic modifications.
Chromosomal locations suitable for genomic integration were identified and characterized to detect possible fitness effects.
We accelerated metabolic engineering by developing a workflow for rapid pathway assembly and pathway optimization in V. natriegens.
Model-predicted parts were used to construct a pathway for maximal 3-hydroxypropionate production in V. natriegens and we demonstrated its functionality.
Via mass spectrometry, we detected our pathway product 3-hydroxypropionic acid in V. natriegens.

B. Marchal