Difference between revisions of "Part:BBa K368000"
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This part consists of two units. First is an inducible promoter (BBa_I0500). The second is a translational fusion of two proteins (pduD-aurF). PduD is a subunit of a diol dehydratase, which is located in the bacterial microcompartment (BMC) of ''Citrobacter freundii'' (Parsons et al, 2010). AurF is a para-aminobenzoate N-oxygenase from ''Streptomyces thioluteus'' (Simurdiak et al., 2006). When pduD was amplified a consensus RBS (AGGAGG) was introduced by using a corresponding primer (GCA GAATTC GCGGCCGC T TCTAGA AGGAGG GTAGTC ATGGAAATCAATGAAAAGC). The two sequences were fused by PCR. For this purpose aurF were amplified with a 5' overhang, which is identical to the last 25 bases of pduD. To obtain a translational fusion this overhang-sequence doesn´t contain the stop codon of pduD. | This part consists of two units. First is an inducible promoter (BBa_I0500). The second is a translational fusion of two proteins (pduD-aurF). PduD is a subunit of a diol dehydratase, which is located in the bacterial microcompartment (BMC) of ''Citrobacter freundii'' (Parsons et al, 2010). AurF is a para-aminobenzoate N-oxygenase from ''Streptomyces thioluteus'' (Simurdiak et al., 2006). When pduD was amplified a consensus RBS (AGGAGG) was introduced by using a corresponding primer (GCA GAATTC GCGGCCGC T TCTAGA AGGAGG GTAGTC ATGGAAATCAATGAAAAGC). The two sequences were fused by PCR. For this purpose aurF were amplified with a 5' overhang, which is identical to the last 25 bases of pduD. To obtain a translational fusion this overhang-sequence doesn´t contain the stop codon of pduD. | ||
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Parsons, J. B., S. Frank, D. Bhella, M. Liang, M. B. Prentice, D. P. Mulvihill & M. J. Warren, (2010) Synthesis of empty bacterial microcompartments, directed organelle protein incorporation, and evidence of filament-associated organelle movement. Mol Cell '''38''': 305-315. | Parsons, J. B., S. Frank, D. Bhella, M. Liang, M. B. Prentice, D. P. Mulvihill & M. J. Warren, (2010) Synthesis of empty bacterial microcompartments, directed organelle protein incorporation, and evidence of filament-associated organelle movement. Mol Cell '''38''': 305-315. | ||
Simurdiak, M., J. Lee & H. Zhao, (2006) A new class of arylamine oxygenases: evidence that p-aminobenzoate N-oxygenase (AurF) is a di-iron enzyme and further mechanistic studies. Chembiochem '''7''': 1169-1172. | Simurdiak, M., J. Lee & H. Zhao, (2006) A new class of arylamine oxygenases: evidence that p-aminobenzoate N-oxygenase (AurF) is a di-iron enzyme and further mechanistic studies. Chembiochem '''7''': 1169-1172. | ||
Revision as of 11:01, 31 July 2010
This part consists of two units. First is an inducible promoter (BBa_I0500). The second is a translational fusion of two proteins (pduD-aurF). PduD is a subunit of a diol dehydratase, which is located in the bacterial microcompartment (BMC) of Citrobacter freundii (Parsons et al, 2010). AurF is a para-aminobenzoate N-oxygenase from Streptomyces thioluteus (Simurdiak et al., 2006). When pduD was amplified a consensus RBS (AGGAGG) was introduced by using a corresponding primer (GCA GAATTC GCGGCCGC T TCTAGA AGGAGG GTAGTC ATGGAAATCAATGAAAAGC). The two sequences were fused by PCR. For this purpose aurF were amplified with a 5' overhang, which is identical to the last 25 bases of pduD. To obtain a translational fusion this overhang-sequence doesn´t contain the stop codon of pduD.
Parsons, J. B., S. Frank, D. Bhella, M. Liang, M. B. Prentice, D. P. Mulvihill & M. J. Warren, (2010) Synthesis of empty bacterial microcompartments, directed organelle protein incorporation, and evidence of filament-associated organelle movement. Mol Cell 38: 305-315.
Simurdiak, M., J. Lee & H. Zhao, (2006) A new class of arylamine oxygenases: evidence that p-aminobenzoate N-oxygenase (AurF) is a di-iron enzyme and further mechanistic studies. Chembiochem 7: 1169-1172.