BBa_K1355004
1
BBa_K1355004
Mercury ions bioremediating device
2014-10-05T11:00:00Z
2015-05-08T01:10:02Z
BBa_K1355001, BBa_K1355000
The biorremediator device is composed by our ???essential biobrick??? (BBa_K1355001) + a MerA gene (BBa_K1355000). In the presence of mercury, The MerR protein will bind to it and MerT, MerP and MerA protein will be expressed and bioremediation will start! The MerP and MerT proteins are responsible for the transport of mercury from the periplasm to the cytoplasm, leading him to the mercury ion reductase (MerA)! The beautiful MerA be responsible for the reduction from Hg2+ in Hg0, which is volatile at room temperature and able to passively leave out the bacteria.
false
false
_1731_
0
15499
9
In stock
true
For this construct, we followed these steps: Extraction of plasmid DNA and DNA quantification of bacteria transformed with the Essential Biobrick (BBa_K1355001) and the bacteria transformed with the MerA Biobrick (BBa_E0840); Verifying the electrophoretic profile of the extracted plasmid DNA; Restriction enzyme digestion for Essential Biobrick with SpeI and PstI and for merA Biobrick with PstI and XbaI; Checking the electrophoretic profile of digested samples; Fragment of interest purification from the gel; Ligation the gel purification of merA gene and digested Essential Biobrick; Transformation of ligation in DH5-alpha; Plasmid DNA extraction from bacteria transformed; Check the electrophoretic profile to see results of samples linked (no fragments); Digestion of Essential Biobrick + merA Biobrick with EcoRI and PstI, aiming to analyze the fragment size to be isolated; Checking the electrophoretic profile of the digested sample to obtain results showing that the isolated fragment is the junction of Essential Biobrick +merA Biobrick ;
false
Luna Barroco de Lacerda, Maria Clara Tavares Astolfi
component2415200
1
BBa_K1355000
component2415196
1
BBa_K1355001
annotation2415200
1
BBa_K1355000
range2415200
1
1198
2975
annotation2415196
1
BBa_K1355001
range2415196
1
1
1189
BBa_K1355000
1
BBa_K1355000
Strong RBS + merA (mercuric ion reductase)+ terminator (BBa_ B0015)
2014-10-05T11:00:00Z
2016-01-13T10:34:49Z
MerA gene sequence is found in the O26-CRL plasmid from Escherichia coli O26. We also added strong RBS from protein 10 found in phage 17 and the double terminator (BBa_B0015) to ensure efficient termination of transcription and messenger RNA recognition.
The MerA gene is one of the most important gene into bacterial mercury-resistance operon. Transforms mercury Hg2+ in Hg0, which is volatile and moves passively out of the bacteria in gaseous form as shown in Figure above. The reduction of Hg2+ occurs by a NADPH-dependent system.
false
false
_1731_
4206
15499
9
In stock
true
We analyzed all Mercury-resistance (Mer) operon genes, in these following strains available at NCBI: (1) Escherichia coli 042, (2) Escherichia coli O26 , (3) Escherichia coli ACN001, (4) Escherichia coli 838C-R1, (5) Escherichia coli APEC-O103, (6) Escherichia coli UMNK88, (7) Escherichia coli AR060302, (8) Escherichia coli PMV-1. It is showed that the mer genes from strains (1) - (2) - (3) - (4) - (5) is conserved among them. However strains (6) - (7) and (8) did not show any similarity. Therefore we used sequences from (1) to (5) due the consensus in E.coli.
false
Luna Barroco de Lacerda
annotation2396893
1
MerA
range2396893
1
24
1639
annotation2396890
1
rbs
range2396890
1
9
16
annotation2396891
1
BBa_ B0015
range2396891
1
1652
1778
BBa_K1355001
1
BBa_K1355001
Regulation and transport of mercury ions
2014-10-05T11:00:00Z
2015-07-23T04:05:30Z
This construction is based on sequences present in the O26-CRL plasmid found in Escherichia coli O26. We added a reverse tryptophan operon terminator to ensure transcription termination of the merR messenger RNA. However we didn???t add transcription terminator after the merP gene, aiming connect this biobrick with others, enabling various functions related to mercury
We call this part an ???Essential biobrick??? designed to be the key piece of any genetic construction related to mercury, targeting both a biosensor as a bioaccumulative. This biobrick has bidirectional promoter, the regulator MerR with a terminator reverse from the tryptophan operon and a forward MerP and MerT transporters.
false
false
_1731_
4206
15499
9
In stock
true
If is desirable to accumulate mercury, assemble this biobrick with a good RBS, a coding sequence of a peptide having affinity to metal and a terminator! Its function will be regulated by presence of mercury through MerR regulator. The bacteria will carry the mercury to cytoplasm until it is linked to the peptide having affinity to the metal, making a inactive mercury. Is not it amazing? This is the strategy that we use in our project! The so called ???Essential Biobrick??? was connected to biobrick for bioremediation, bioaccumulation and bio-detection!
false
Luna Barroco de Lacerda
annotation2415134
1
Bidirecional promoter
range2415134
1
510
538
annotation2415127
1
Bidirecional promoter
range2415127
1
486
514
annotation2415143
1
RBS
range2415143
1
901
907
annotation2415126
1
MerR
range2415126
1
44
480
annotation2415155
1
MerP
range2415155
1
914
1189
annotation2415752
1
RBS
range2415752
1
537
543
annotation2415141
1
MerT
range2415141
1
551
900
annotation2415749
1
RBS
range2415749
1
490
496
annotation2397153
1
Trp Terminator
range2397153
1
1
43
BBa_K1355004_sequence
1
aaagaaagttaaaatgccgccagcggaactggcggctgtgggactaaggcatagctgaccttgccaggcctgcttcgccctgtagtgacgcgatcaacgggcaggaaacattcccctttcgtgcatggcaggcgcacacgagttcagacagcacggtttccatgcgcgccaagtcggccatcttctcgcgcacgtccttgagcttgtgttcggccaggctgctggcctcctcgcagtgggtgccatcgtcgagccgcaacagctcggcaatctcgtccagactgaaccccagccgctgtgccgatttcacgaatttcacccgaaccacgtccgcctccccatagcggcggatgctgccgtaaggcttgtccggttcccgcaacaggcccttgcgctgatagaagcggattgtctccacgttgaccccggccgccttggcaaaaacgccaatggtcaggttttccaaattattttccatatcgcttgactccgtacatgagtacggaagtaaggttacgctatccaatccaaattcaaaagggccaacgtatgtctgaaccacaaaacgggcgcggtgcgctcttcgccggcgggctggccgccattcttgcatcgacctgctgcctggggccgctagtactggtcgccctgggcttctccggtgcttggatcggcaacctgacggtgctggaaccctatcgaccgttgttcatcggcgcggcgctagtggcgctgttcttcgcctggaagcggatttaccggcccgtgcaggcatgcaagccaggtgaggtctgcgcgattccgcaggtgcgcgccacctacaagctgattttctggatcgtggccgtgctggtcctggtcgcgcttggatttccctatgtcgttccatttttctattaaccaggagttcatcatgaagaaactgtttgcctcccttgccctcgccgccgctgttgccccggtgtgggccgctacccagaccgtcacgctagcggttcccggcatgacttgcgccgcctgcccgatcacagtcaagaaagcgctctccaaggtcgaaggcgtgagcaaggtcgatgtgggcttcgagaagcgcgaggccgtcgtcacttttgacgacaccaaggccagcgtacagaagctgaccaaggccaccgcagacgccggctatccgtccagcgtcaagcagtgatactagagaactttaagaaggagatatatccatgcaatcagcggatgtctcctacgccaagggcagcgccaagctcgccattgaggtcggcacgtcacccgacgcgctgacggccgctgtagctggactcggttatcgggccacgctggccgatgccccctcagtttcgacgccgggcggattgctcgacaagatgcgcgatctgctgggcagaaacgacaagacgggtagcagcggcgcattgcatatcgccgtcatcggcagcggcggggccgcgatggcagcggcgctgaaggccgtcgagcaaggcgcacgtgtcacgctgatcgagcgcggcaccatcggcggcacctgcgtcaatgtcggttgtgtgccgtccaagatcatgatccgcgccgcccatatcgcccatctgcgccgggaaagcccgttcgatggcggcatcgccgctaccacgccgaccatccagcgcacggcgctgctggcccagcagcaggcccgcgtcgatgaactgcgccacgccaagtacgaaggcatcttggagggcaatccggcgatcactgtgctgcacggctccgcccgctttaaggacaatcgcaacctgatcgtgcaactcaacgacggcggcgagcgcgtggtggcattcgaccgctgcctgatcgccaccggcgcgagcccggccgtgccgccgattcccggcctgaaagacactccgtactggacttccactgaagcgctggtcagcgagacgattcctaagcgcctggccgtgattggctcatcagtggtggcgctggagctggcgcaggcgttcgcccgactcggagcgaaggtgacgatcctggctcgcagcacgctgttcttccgcgaagacccagctataggcgaagccgtcacggccgcattccgcatggagggcatcgaggtgagggaacacacccaggccagccaggtcgcgtatatcaatggtgaaggggacggcgagttcgtgctcaccacggcgcacggcgaactgcgcgccgacaagctgctggtcgccaccggccgcgcgcccaacacacgcaagctggcactggatgcgacgggcgtcacgctcaccccgcaaggcgctatcgtcatcgaccccggcatgcgtacaagcgtggaacacatctacgccgcaggcgactgcaccgaccagccgcagttcgtctatgtggcggcagcggccggcactcgcgccgcgatcaacatgaccggcggtgacgcggccctgaacctgaccgcgatgccggccgtggtgttcaccgacccgcaagtggcgaccgtaggctacagcgaggcggaagcgcaccatgacggcatcaaaactgatagtcgcacgctaacgctggacaacgtgccgcgcgcgctcgccaacttcgacacgcgcggcttcatcaaactggtggttgaagaaggcagcggacgactgatcggcgtgcaggcagtggccccggaagcgggcgaactgatccagacggccgcactggcgattcgcaaccggatgacggtgcaggaactggccgaccagttgttcccctacctgacgatggtcgaagggttgaagctcgcggcgcagaccttcaacaaggatgtgaagcagctttcctgctgcgccgggtgaggactactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata
BBa_K1355000_sequence
1
aactttaagaaggagatatatccatgcaatcagcggatgtctcctacgccaagggcagcgccaagctcgccattgaggtcggcacgtcacccgacgcgctgacggccgctgtagctggactcggttatcgggccacgctggccgatgccccctcagtttcgacgccgggcggattgctcgacaagatgcgcgatctgctgggcagaaacgacaagacgggtagcagcggcgcattgcatatcgccgtcatcggcagcggcggggccgcgatggcagcggcgctgaaggccgtcgagcaaggcgcacgtgtcacgctgatcgagcgcggcaccatcggcggcacctgcgtcaatgtcggttgtgtgccgtccaagatcatgatccgcgccgcccatatcgcccatctgcgccgggaaagcccgttcgatggcggcatcgccgctaccacgccgaccatccagcgcacggcgctgctggcccagcagcaggcccgcgtcgatgaactgcgccacgccaagtacgaaggcatcttggagggcaatccggcgatcactgtgctgcacggctccgcccgctttaaggacaatcgcaacctgatcgtgcaactcaacgacggcggcgagcgcgtggtggcattcgaccgctgcctgatcgccaccggcgcgagcccggccgtgccgccgattcccggcctgaaagacactccgtactggacttccactgaagcgctggtcagcgagacgattcctaagcgcctggccgtgattggctcatcagtggtggcgctggagctggcgcaggcgttcgcccgactcggagcgaaggtgacgatcctggctcgcagcacgctgttcttccgcgaagacccagctataggcgaagccgtcacggccgcattccgcatggagggcatcgaggtgagggaacacacccaggccagccaggtcgcgtatatcaatggtgaaggggacggcgagttcgtgctcaccacggcgcacggcgaactgcgcgccgacaagctgctggtcgccaccggccgcgcgcccaacacacgcaagctggcactggatgcgacgggcgtcacgctcaccccgcaaggcgctatcgtcatcgaccccggcatgcgtacaagcgtggaacacatctacgccgcaggcgactgcaccgaccagccgcagttcgtctatgtggcggcagcggccggcactcgcgccgcgatcaacatgaccggcggtgacgcggccctgaacctgaccgcgatgccggccgtggtgttcaccgacccgcaagtggcgaccgtaggctacagcgaggcggaagcgcaccatgacggcatcaaaactgatagtcgcacgctaacgctggacaacgtgccgcgcgcgctcgccaacttcgacacgcgcggcttcatcaaactggtggttgaagaaggcagcggacgactgatcggcgtgcaggcagtggccccggaagcgggcgaactgatccagacggccgcactggcgattcgcaaccggatgacggtgcaggaactggccgaccagttgttcccctacctgacgatggtcgaagggttgaagctcgcggcgcagaccttcaacaaggatgtgaagcagctttcctgctgcgccgggtgaggactactagagccaggcatcaaataaaacgaaaggctcagtcgaaagactgggcctttcgttttatctgttgtttgtcggtgaacgctctctactagagtcacactggctcaccttcgggtgggcctttctgcgtttata
BBa_K1355001_sequence
1
aaagaaagttaaaatgccgccagcggaactggcggctgtgggactaaggcatagctgaccttgccaggcctgcttcgccctgtagtgacgcgatcaacgggcaggaaacattcccctttcgtgcatggcaggcgcacacgagttcagacagcacggtttccatgcgcgccaagtcggccatcttctcgcgcacgtccttgagcttgtgttcggccaggctgctggcctcctcgcagtgggtgccatcgtcgagccgcaacagctcggcaatctcgtccagactgaaccccagccgctgtgccgatttcacgaatttcacccgaaccacgtccgcctccccatagcggcggatgctgccgtaaggcttgtccggttcccgcaacaggcccttgcgctgatagaagcggattgtctccacgttgaccccggccgccttggcaaaaacgccaatggtcaggttttccaaattattttccatatcgcttgactccgtacatgagtacggaagtaaggttacgctatccaatccaaattcaaaagggccaacgtatgtctgaaccacaaaacgggcgcggtgcgctcttcgccggcgggctggccgccattcttgcatcgacctgctgcctggggccgctagtactggtcgccctgggcttctccggtgcttggatcggcaacctgacggtgctggaaccctatcgaccgttgttcatcggcgcggcgctagtggcgctgttcttcgcctggaagcggatttaccggcccgtgcaggcatgcaagccaggtgaggtctgcgcgattccgcaggtgcgcgccacctacaagctgattttctggatcgtggccgtgctggtcctggtcgcgcttggatttccctatgtcgttccatttttctattaaccaggagttcatcatgaagaaactgtttgcctcccttgccctcgccgccgctgttgccccggtgtgggccgctacccagaccgtcacgctagcggttcccggcatgacttgcgccgcctgcccgatcacagtcaagaaagcgctctccaaggtcgaaggcgtgagcaaggtcgatgtgggcttcgagaagcgcgaggccgtcgtcacttttgacgacaccaaggccagcgtacagaagctgaccaaggccaccgcagacgccggctatccgtccagcgtcaagcagtga
igem2sbol
1
iGEM to SBOL conversion
Conversion of the iGEM parts registry to SBOL2.1
Chris J. Myers
James Alastair McLaughlin
2017-03-06T15:00:00.000Z