Team:LZU-China/tempfortest

          In wet lab, we designed a novel MFC device containing genetic engineered bacteria. For anode strain, we’ve cloned a PNP sensor sequence and a riboflavin into Escherichia coli. The recombinants is able to detect PNP and produce riboflavin to boost electrical generation when co-cultured with Shewanella oneidensis.

          We found that the part BBa_K381001 by iGEM10_BCCS-Bristol is a sensor which can detect the appearance of nitrate and nitrite. So we perfomed an experience by this part to see if this part can detect PNP(p-Nitrophenol). We got a good result.You can see the PNP can also induce the green fluorescence.

                                                              Figure-1 The pathway of PNP sensor

                                                              Figure-1 The pathway of PNP sensor

                                                              Figure-1 The pathway of PNP sensor

          We found four genes to reduce the Cr(VI) to Cr(III). They are nahG, nahR, nahE, chrR and yieF. We didn’t do the deep research about them because of time limitation. They can be used in the general plasmids such as pBR322. We just made them to be the formation of Biobrick. More uses and functions need to be demonstrated and found.

nahG: We got the sequence from Pseudomonas putida

          (Primer:sense:5’---CCGGAATTCGCGGCCGCTTCTAGATGAAAAACAATAAACCTGGCTTGCGC---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGTCACCCTTGACGTAGCACACC---3’)
nahE: We got the sequence from Pseudomonas putida
          (Primer:sense:5’---CCGGAATTCGCGGCCGCTTCTAGATGTCGAATAAAATTATGAAAACGTCGCG---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGCTACTTCAATTCATTACTGTATTTAGCGTG---3’)
nahR: We got the sequence from Pseudomonas putida
          (Primer:sense:5’--- CCGGAATTCGCGGCCGCTTCTAGATGAAAAACAATAAACCTGGCTTGCGC---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGTCACCCTTGACGTAGCACACC---3’)
chrR: We got the sequence from Escherichia coli str. K-12
          (Primer:sense:5’--- CCGGAATTCGCGGCCGCTTCTAGATGAAAAAAATAGTCCAGTCGGAAGG---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGTCAGGCCTTTTTGTGCTGTTCAAC---3’)
yieF: We got the sequence from Escherichia coli str. K-12
          (Primer:sense:5’--- GAATTCGGATCCATGTCTGAAAAATTGCAGGTGG---3’;
          anti-sense:5’---GAATTCTCTAGATTAGATCTTAACTCGCTGAATAAA---3’)

 

     (Primer:sense:5’--- CCGGAATTCGCGGCCGCTTCTAGATGAAAAAAATAGTCCAGTCGGAAGG---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGTCAGGCCTTTTTGTGCTGTTCAAC---3’)
yieF: We got the sequence from Escherichia coli str. K-12
          (Primer:sense:5’--- GAATTCGGATCCATGTCTGAAAAATTGCAGGTGG---3’;
          anti-sense:5’---GAATTCTCTAGATTAGATCTTAACTCGCTGAATAAA---3’)

 

     (Primer:sense:5’--- CCGGAATTCGCGGCCGCTTCTAGATGAAAAAAATAGTCCAGTCGGAAGG---3’;
          anti-sense:5’---AAAATACTAGTAGCGGCCGCTGCAGTCAGGCCTTTTTGTGCTGTTCAAC---3’)
yieF: We got the sequence from Escherichia coli str. K-12
          (Primer:sense:5’--- GAATTCGGATCCATGTCTGAAAAATTGCAGGTGG---3’;
          anti-sense:5’---GAATTCTCTAGATTAGATCTTAACTCGCTGAATAAA---3’)