Our aim is to modify the unicellular microalga <i>Chlamydomonas reinhardtii</i> for effective trinitrotoluene (TNT)<br> biodegradation by expression of Enterobacter cloacae nitroreductase gene nfsI. <i>C. reinhardtii</i> is a ubiquitous <br>species capable of thriving in soil, freshwater and marine environments, making this alga a suitable choice for<br> removal of TNT from a wide variety of biomes. We have opted for the use of nfsI as the gene in question is<br> a well-characterized nitroreductase with a known sequence. Our experimental procedure involves (a) development<br> of a synthetic nfsI gene with flanking prefix and suffixes of standard Biobricks, (b) ligation of this sequence<br> to the C. heinhardtii expression vector pRbcBRL, (c) transfection of <i>C. reinhardtii</i> with the recombinant plasmid<br> and (d) TNT tolerance and biodegradation capacity assessment of the modified alga. <br>
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Our aim is to modify the unicellular microalga <i>Chlamydomonas reinhardtii</i> for effective trinitrotoluene (TNT) biodegradation by expression of Enterobacter cloacae nitroreductase gene nfsI. <i>C. reinhardtii</i> is a ubiquitous species capable of thriving in soil, freshwater and marine environments, making this alga a suitable choice for removal of TNT from a wide variety of biomes. We have opted for the use of nfsI as the gene in question is a well-characterized nitroreductase with a known sequence. Our experimental procedure involves (a) development of a synthetic nfsI gene with flanking prefix and suffixes of standard Biobricks, (b) ligation of this sequence to the C. heinhardtii expression vector pRbcBRL, (c) transfection of <i>C. reinhardtii</i> with the recombinant plasmid and (d) TNT tolerance and biodegradation capacity assessment of the modified alga.
Biodegradation of TNT and TNT derivatives by nfsI-transfected Chlamydomonas Reinhardtii
Abstract
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Our aim is to modify the unicellular microalga Chlamydomonas reinhardtii for effective trinitrotoluene (TNT) biodegradation by expression of Enterobacter cloacae nitroreductase gene nfsI. C. reinhardtii is a ubiquitous species capable of thriving in soil, freshwater and marine environments, making this alga a suitable choice for removal of TNT from a wide variety of biomes. We have opted for the use of nfsI as the gene in question is a well-characterized nitroreductase with a known sequence. Our experimental procedure involves (a) development of a synthetic nfsI gene with flanking prefix and suffixes of standard Biobricks, (b) ligation of this sequence to the C. heinhardtii expression vector pRbcBRL, (c) transfection of C. reinhardtii with the recombinant plasmid and (d) TNT tolerance and biodegradation capacity assessment of the modified alga.
Something about spirit of being team
In this summer; we learned about iGEM, how it works, what the biggest part of competition is and how we can win a medallion.
When we were newbie, we organized a lot of meeting to find out our main project than we got idea for improving human welfare so we choose an idea from brainstorm results. We searched to get a gene to work on. As a result; lab works began.
We added our protocols which are helpful to transform Chlamydomonas reinhardtii and Escherichia coli. Basic cloning protocols are available on our protocol page. Experiments and safety questions also available from link at the top section.
IGEM Project is our number one priority. As we worked really hard for our project it is important for us to make our dreams come true which is taking a gold medal. Questioning different items like experiment time, materials and methods and the efficiency of the experiments improved our skills and we started to look as a scientist to problems we faced. There were lots of challenging parts and sometimes we thought to quit but we listened to “Eye of The Tiger” and we did not give up from our experiments we delayed them yes, but we never gave up.
IGEM team- meet-up was a great opportunity for us to communicate with the other groups, understood their projects and share our ideas. As we had a chance for an interview with Anatolia Agency, we also used media power to get attention to our project, iGEM and the importance of synthetic biology.
Apart from that we did not forget having fun apart from our lab. As we planned a BBQ party, it was also a great chance for us to relax and gain some energy for our experiments, I did not even mention about the delicious foods and how joyful was our activities. Also like in iGEM team meeting up, we had a chance to introduce our project and main aim to other scientists and researchers in different areas.
Links at above lead you to specific event and its photos suppose to be at that link. If not, look at gallery link.
Thanks and get great time.
Favorite Parts
BBa_K596001
BBa_K596004
DNA Submission to Registry
bunu koyun yanlis olmus https://2011.igem.org/File:DNA_submission_to_registry.JPG
We are really grateful to Gulce Itir Percin, Aydan Torun,Ömer Faruk Sarıoglu and Zeynep E.
Ulger for their help and especially guidance of Mrs. Ulger about unknown
lab equipments. Prof. Dr. Tayfun Özçelik and Assistant Prof. Ali Güre are helped us to start iGEM team. Assistant Prof. Ayşe Tekinay and her lab were assisted our project a lot. Also we thanks to other members of SBL and NBT for patience.
Sponsors
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