The TAL1 is a TALEN which came from a plant pathogenic bacteria in the genus Xanthomonas.
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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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.
Revision as of 12:19, 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.
