Difference between revisions of "Part:BBa K2915225"

 
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<partinfo>BBa_K2915225 short</partinfo>
 
<partinfo>BBa_K2915225 short</partinfo>
  
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The Talen 1 it's a Talen which came from a plant pathogenic bacteria in the genus Xanthomonas. These designed proteins can able to bind specific DNA fragment ( 5'CTGATC3') such as the human amylase gene . This TAL1 can be purify because of the presence of a 6His-Tag in the N-terminal of the protein.
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The TAL1 is a TALEN which came from a plant pathogenic bacteria in the genus Xanthomonas.
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The promoter T7 and RBS (Bba_K525998) are regulator that are assembled with TAL1, which allows the production TAL1 with its HisTag using E.coli DH5-alpha bacteria. This part can bind itself with a DNA sequence of 5’ CTGATC 3’ (BM1). This engineered TAL consist of repeat domain has a primary structure with two amino acids that differs in each domain, those two amino acids are a code for one nucleotide of the binding sequence selected. HD, NG, NN, NI, NG and HD are the two amino that are changed in the CTGATC sequence, respectfully.
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'''Usage and Biology'''
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This Transcription Activator-Like (TAL) is a specific DNA binding protein, that is designed to recognize a specific DNA binding motif (BM1). This BM1 are present on the DNA that are amplify by PSR.
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'''Production'''
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To verify the production of TAL1, an SDS PAGE was performed and stained with Coomassie blue
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Different growing conditions were tested to determine the best growing conditions. The production was also tested two types E.coli bacteria :
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-DH5a
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-BL21 DE3
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(0)
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Figure 1. SDS-PAGE of the production in DH5a of TAL, well 1, 2 and 3 are not induced of TAL1, TAL2 clone1 and TAL2 clone2, respectively. Well 4, 5 and 6 are induced at 30°C with 0.1M IPTG for 3 hours of TAL1, TAL2 clone1 and TAL2 clone2, respectively.  
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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here

Revision as of 12:14, 18 October 2019


IPTG inducible promoter (T7) with RBS TAL1 6-His-Tag with TEV site

The TAL1 is a TALEN which came from a plant pathogenic bacteria in the genus Xanthomonas. The promoter T7 and RBS (Bba_K525998) are regulator that are assembled with TAL1, which allows the production TAL1 with its HisTag using E.coli DH5-alpha bacteria. This part can bind itself with a DNA sequence of 5’ CTGATC 3’ (BM1). This engineered TAL consist of repeat domain has a primary structure with two amino acids that differs in each domain, those two amino acids are a code for one nucleotide of the binding sequence selected. HD, NG, NN, NI, NG and HD are the two amino that are changed in the CTGATC sequence, respectfully.

Usage and Biology This Transcription Activator-Like (TAL) is a specific DNA binding protein, that is designed to recognize a specific DNA binding motif (BM1). This BM1 are present on the DNA that are amplify by PSR.

Production To verify the production of TAL1, an SDS PAGE was performed and stained with Coomassie blue Different growing conditions were tested to determine the best growing conditions. The production was also tested two types E.coli bacteria : -DH5a -BL21 DE3 (0)

Figure 1. SDS-PAGE of the production in DH5a of TAL, well 1, 2 and 3 are not induced of TAL1, TAL2 clone1 and TAL2 clone2, respectively. Well 4, 5 and 6 are induced at 30°C with 0.1M IPTG for 3 hours of TAL1, TAL2 clone1 and TAL2 clone2, respectively.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]