Difference between revisions of "Part:BBa K562009"

Revision as of 13:41, 21 September 2011

Salty_PduABTUNJK BMC Components

This is a composite part comprising a constitutive promoter, which is the tatABCD promoter from E. coli K-12, driving production of the PduA, PduB, PduT, PduU and PduN proteins from Salmonella enterica serovar Typhimurium LT2 (identical to part BBa_K562008) AND proteins PduJ and PduK. These are all components of the Salmonella bacterial microcompartment (BMC). The PduB, PduU, PduN and PduK gene products all carry C-terminal 6-His affinity/epitope tags. Production of the PduB, PduU, PduN and PduK proteins has been verified by Western immunoblotting (anti-penta-His) and production of PduA, PduB, PduT, PduU, PduN, PduJ and PduK has been confirmed by 35S-Met labelling. The proteins have also been purified from E. coli by immobilised metal affinity chromatography (IMAC) and identified by tryptic peptide mass fingerprinting. The construct is cloned as an EcoRI / PstI fragment into pSB1C3. The clone is also known as pSB-ABTUNJK in the Sargent Laboratory, Dundee, UK.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 1898
Illegal BamHI site found at 3058
Illegal BamHI site found at 3197
Illegal BamHI site found at 3291
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal NgoMIV site found at 1441
Illegal NgoMIV site found at 2685
Illegal AgeI site found at 1426
Illegal AgeI site found at 2040
Illegal AgeI site found at 2952
Illegal AgeI site found at 2988
1000
COMPATIBLE WITH RFC[1000]

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 <partinfo>BBa_K562009 short</partinfo>
-This is a composite part comprising a constitutive promoter, which is the <i>tatABCD</i> promoter from <i>E. coli</i> K-12, driving production of the PduA, PduB, PduT, PduU and PduN proteins from Salmonella enterica serovar Typhimurium LT2 (identical to part BBa_K562008) AND proteins PduJ and PduK. These are all components of the <i>Salmonella</i> bacterial microcompartment (BMC). The PduB, PduU, PduN and PduK gene products all carry C-terminal 6-His affinity/epitope tags. Production of the PduB, PduU, PduN and PduK proteins has been verified by Western immunoblotting (anti-penta-His) and production of PduA, PduB, PduT, PduU, PduN, PduJ and PduK has been confirmed by 35S-Met labelling. The proteins have also been purified from E. coli by immobilised metal affinity chromatography (IMAC) and identified by tryptic peptide mass fingerprinting. The construct is cloned as an EcoRI / PstI fragment into pSB1C3. The clone is also known as pSB-ABTUNJK in the Sargent Laboratory, Dundee, UK.
+This is a composite part comprising a constitutive promoter, which is the <i>tatABCD</i> promoter from <i>E. coli</i> K-12, driving production of the PduA, PduB, PduT, PduU and PduN proteins from Salmonella enterica serovar Typhimurium LT2 (identical to part BBa_K562008) AND proteins PduJ and PduK. These are all components of the <i>Salmonella</i> bacterial microcompartment (BMC). The PduB, PduU, PduN and PduK gene products all carry C-terminal 6-His affinity/epitope tags. Production of the PduB, PduU, PduN and PduK proteins has been verified by Western immunoblotting (anti-penta-His) and production of PduA, PduB, PduT, PduU, PduN, PduJ and PduK has been confirmed by 35S-Met labelling. The proteins have also been purified from <i>E. coli</i> by immobilised metal affinity chromatography (IMAC) and identified by tryptic peptide mass fingerprinting. The construct is cloned as an EcoRI / PstI fragment into pSB1C3. The clone is also known as pSB-ABTUNJK in the Sargent Laboratory, Dundee, UK.
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