Stanford/BIOE44:Module3:Day2:nitrate II: Difference between revisions

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'''Nitrate Sensor Project'''
'''Nitrate Sensor Project II'''


Authors: Daniel Bui and Aaditya Shidham
Authors: Daniel Bui and Aaditya Shidham
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Created BioBrick Parts: BBa_fnr001, BBa_narGp001
Created BioBrick Parts: BBa_fnr001, BBa_narGp001


Long Description: We plan to develop a system that can sense a low level of nitrate in water by using a mutant of nitrate-inducible fnr coupled with the narG promoter. While normally only inducible under anaerobic conditions, a single silent mutation to convert leucine-28 to histidine can allow the fnr gene to operate under anaerobic conditions. The fnr gene acts as a promoter for narG, which can send a PoPs-out signal to a GFP reporter. Previous studies have shown that the mutated form of fnr can sense nitrate at as low as 0.1 uM.
Long Description: We plan to develop a system that can sense a low level of nitrate in water by using a mutant of nitrate-inducible fnr coupled with the narG promoter. While normally only inducible under anaerobic conditions, a single silent mutation to convert leucine-28 to histidine can allow the fnr gene to operate under aerobic conditions. The fnr gene acts as a promoter for narG, which can send a PoPs-out signal to a GFP reporter. Previous studies have shown that the mutated form of fnr can sense nitrate at as low as 0.1 uM.


''BBa_narGp001 sequence:''
''BBa_narGp001 sequence:''

Revision as of 22:20, 4 May 2010

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Nitrate Sensor Project II

Authors: Daniel Bui and Aaditya Shidham

Created BioBrick Parts: BBa_fnr001, BBa_narGp001

Long Description: We plan to develop a system that can sense a low level of nitrate in water by using a mutant of nitrate-inducible fnr coupled with the narG promoter. While normally only inducible under anaerobic conditions, a single silent mutation to convert leucine-28 to histidine can allow the fnr gene to operate under aerobic conditions. The fnr gene acts as a promoter for narG, which can send a PoPs-out signal to a GFP reporter. Previous studies have shown that the mutated form of fnr can sense nitrate at as low as 0.1 uM.

BBa_narGp001 sequence:

GAATTCGCGGCCGCTTCTAGAGATCCTAAAGGGGTATCTTAGGAATTTACTTTATTTTTCATCCCC ATCACTCTTGATCGTTATCAATTCCCACGCTGTTTCAGAGCGTTACCTTGCCCTTAAACATTAGCA ATGTCGATTTATCAGAGGGCCGACAGGCTCCCACTACTAGTAGCGGCCGCTGCAG

The promoter itself does not contain any BioBrick restriction enzyme sites, so this part is in accordance with BioBrick Standard 10.

fnr operator/promoter sequence: GAATTC GCGGCCGC T TCTAGAG TATATTTACATCTAATATCGAAATTCTCTGCTGTTAAGGTTTGCTTAGACTTAC TTGCTCCCTAAAAAGATGTTAAAATTGACAAATATCAATTACGGCTT T ACTAGT A GCGGCCG CTGCAG

fnr ORF sequence: GAATTC GCGGCCGC T TCTAG ATGATCCCGGAAAAGCGAATTATACGGCGCATTCAGTCTGGCGGTTGTGCTATCCATTGCCAGGATTGC AGCATCAGCCAGCATTGCATCCCGTTCACACTCAACGAACATGAGCTTGATCAGCTTGATAATATCATT GAGCGGAAGAAGCCTATTCAGAAAGGCCAGACGCTGTTTAAGGCTGGTGATGAACTTAAATCGCTTTAT GCCATCCGCTCCGGTACGATTAAAAGTTATACCATCACTGAGCAAGGCGACGAGCAAATCACTGGTTTC CATTTAGCAGGCGACCTGGTGGGATTTGACGCCATCGGCAGCGGCCATCACCCGAGCTTCGCGCAGGCG CTGGAAACCTCGATGGTATGTGAAATCCCGTTCGAAACGCTGGACGATTTGTCCGGTAAAATGCCGAAT CTGCGTCAGCAGATGATGCGTCTGATGAGCGGTGAAATCAAAGGCGATCAGGACATGATCCTGCTGTTG TCGAAGAAAAATGCCGAGGAACGTCTGGCTGCATTCATCTACAACCTGTCCCGTCGTTTTGCCCAACGC GGCTTCTCCCCTCGTGAGTTCCGCCTGACGATGACTCGTGGCGATATCGGTAACTATCTGGGCCTGACG GTAGAAACCATCAGCCGTCTGCTGGGTCGCTTCCAGAAAAGCGGCATGCTGGCAGTCAAAGGTAAATAC ATCACCATCGAAAATAACGATGCGCTGGCCCAGCTTGCTGGTCATACGCGTAACGTTGCCTAATAA T ACTAGT A GCGGCCG CTGCAG