Team:IIT Delhi/results

Parts Submitted

Part:BBa_K1866000
Promoter+RibosomeBindingSite+NrfA
The assimilatory Nitrite Reductase gene under the control of a strong Promoter Anderson J23104 with a Ribosome Binding Site was cloned onto the pSB1A3 plasmid backbone. This gene catalyses the conversion of Nitrite (NO2 -) to Ammonium (NH4 +).

Part:BBa_K1866001
NosZ gene+SuperYellowFluorescenceProtein
This part consists of NosZ gene and a sequence encoding for Super Yellow Fluorescence Protein downstream to it. NosZ gene product catalyses the conversion of Nitrous Oxide (N2O) to molecular Nitrogen gas in a process called denitrification.

Part:BBa_K1866002
Promoter+RibosomeBindingSite+NrfA+SYFP
This part has the Super Yellow Fluorescence Protein tagged to the NrfA gene product from our biobrick BBa_K186600

Part:BBa_K1866003
Promoter+RibosomeBindingSite+Sqr gene
This part consists of the Sulfide Quinone Reductase Sqr gene under the control of a strong promoter with a Ribosome Binding Site. The Sqr gene catalyses the conversion of hydrogen Sulfide gas to elemental Sulfur.

Part:BBa_K1866004
Promoter+RibosomeBindingSite+NosZ+SuperYellowFluorescenceProtein
This part consists of the gene NosZ that encodes for an enzyme named Dissimilatory Nitrous Oxide Reductase. This enzyme catalyses the conversion of Nitrous Oxide, a greenhouse gas to Nitrogen gas. The NosZ gene is cloned under the control of a strong Promoter with a Ribosome Binding Site for optimum expression. This part also consists of a Super Yellow Fluorescence Protein tag to check the expression of the NosZ gene.

Part:BBa_K1866006
Promoter+RibosomeBindingSite+NrfA (in pSB1C3)
The assimilatory Nitrite Reductase gene under the control of a strong Promoter Anderson J23104 with a Ribosome Binding Site was cloned onto the pSB1C3 plasmid backbone. This gene catalyses the conversion of Nitrite (NO2 -) to Ammonium (NH4 +).

Part:BBa_K1866007
NosZ+SuperYellowFluorescenceProtein (in pSB1C3)
This part consists of NosZ gene and a sequence encoding for Super Yellow Fluorescence Protein downstream to it. NosZ gene product catalyses the conversion of Nitrous Oxide (N2O) to molecular Nitrogen gas in a process called denitrification. This part can be cloned under the control of a promoter of choice and its expression can be readily analyzed by checking for the expression of the yellow fluorescence protein.

Part:BBa_K1866008
Promoter+RibosomeBindingSite+NrfA+SuperYellowFluorescenceProtein (in pSB1C3)
This part has the Super Yellow Fluorescence Protein tagged to the NrfA gene product. The Promoter+RBS+NrfA construct was obtained from our biobrick BBa_K1866000. The RBS+SYFP construct was procured from the biobrick BBa_K864101.

Part:BBa_K1866009
promoter+RibosomeBindingSite+Sqr (in pSB1C3)
This part consists of the Sulfide Quinone Reductase Sqr gene under the control of a strong promoter with a Ribosome Binding Site. The Sqr gene catalyses the conversion of hydrogen Sulfide gas to elemental Sulfur.

Part:BBa_K1866010
NosZ gene+SuperYellowFluorescenceProtein (in pSB1A3)
This part consists of NosZ gene and a sequence encoding for Super Yellow Fluorescence Protein downstream to it. NosZ gene product catalyses the conversion of Nitrous Oxide (N2O) to molecular Nitrogen gas in a process called denitrification. This part can be cloned under the control of a promoter of choice and its expression can be readily analyzed by checking for the expression of the yellow fluorescence protein.

Clone Confirmation Using Agarose Gel Electrophoresis

final gel.jpg
Characterization Using SDS PAGE



Characterization Using Indophenol Test



Characterization Using pH Monitoring



Characterization Using Nessler’s test







Criteria Fulfilled


For Bronze:
  • We registered for iGEM, and are going to attend the Giant Jamboree.
  • We have completed the Judging Form.
  • We have created and shared a description of the team's project using the iGEM wiki, and documented the team's parts using the Registry of Standard Biological Parts.
  • We are going to present a poster and a talk at the iGEM Jamboree.
  • We have created a page on our team wiki with a clear attribution of each aspect of our project, following all the necessary criteria as per the instructions.
  • We have document a new standard BioBrick Part, which is central to our project and are going to submit this part to the iGEM Registry, adhering to the iGEM Registry guidelines.


    • For Silver:
  • We have experimentally validated that our BioBrick Part as well as the device (of our own design and construction) works as per the expectations. Apart from that, we also documented the characterization of this part in the Main Page section of the Registry.
  • We submitted this new part to the iGEM Parts Registry.
  • Our team has identified the issue of Pollution in general. We have investigated the effect it has through numerous surveys carried out in different parts of Delhi. We are implementing our prototype in project "Aanch" of Enactus. And finally, we have published a documentation on Pollution to make people aware of the seriousness of the issue.


    • For Gold:
  • We are mentoring the iGEM IIT Kharagpur team, participating for the first time in the Giant Jamboree Competition.
  • We have improved the characterization of our previously existing BioBrick Part.
  • We have designed a functional prototype of our project.
  • And finally, we integrated the issues raised during the surveys, through necessary changes in our design.

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    Acknowledgement

    A few months back, it was a dream. Today, it’s a reality. The transition didn't come by our efforts alone. There were people who didn’t hesitate to spend their valuable time on us. And we certainly do owe a lot to them. We are grateful to our mentor Prof. Shaikh Ziauddin Ahammad, who selflessly supported us in unfavorable situation, for facilitating funding and allowing us access to UG lab. It was a fortunate coincidence that Dr. Stefan Oehler accepted role of primary trouble shooter for our molecular biology work, and happily answered our queries. We are grateful to him that he allowed us to leach his lab resources. We are also thankful to Prof. James Gomes, from the Kusuma School of Biological Sciences, for his valuable advice and moral support, and Prof. Biswajit Kundu for helpful conversations.

    A special note is due to Prof. P.V.M Rao who went beyond his way to facilitate our work and for letting us the access to Workshop, IIT Delhi. We are thankful to Prof. Anurag Sharma, Dean, IIT Delhi for helping us sail administrative processes. Our team owes a lot to Anamika Singh (RNA 2 Lab), who provided constant support and help, no matter how late it was in the night. Also a thank you note to Garima Goyal (Protein Engineering Lab) for allowing us to access to her labs. We are also thankful to the Karan Varshney for helping us make crowd funding video.