Difference between revisions of "Part:BBa K5291032"
 
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As a plasmid, pBBR1MCS2-<i>nadM-nadK-pntA-pntB</i> can increase the intracellular level of NADPH by introducing four exogenous genes: <i>nadM</i>, <i>nadK</i>, <i>pntA</i> and <i>pntB</i>.
 
As a plasmid, pBBR1MCS2-<i>nadM-nadK-pntA-pntB</i> can increase the intracellular level of NADPH by introducing four exogenous genes: <i>nadM</i>, <i>nadK</i>, <i>pntA</i> and <i>pntB</i>.
  
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===Usage and Biology===
 
===Usage and Biology===
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To increase the intracellular level of NADPH, we constructed the pBBR1MCS2-<i>nadM-nadK-pntA-pntB</i> plasmid containing four exogenous genes, including nadM, nadK, pntA, and pntB.<br>
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<html><img width = "600" src="https://static.igem.wiki/teams/5291/images/parts-lhs/pbbr1mcs2-pnta-pntb-nadm-nadk.png" /></html><br>
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<b>Fig.1 The map of plasmid pBBR1MCS2-<i>nadM-nadK-pntA-pntB</i>.</b><br><br>
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Since the efficiency of homologous recombination of multiple fragments was too low, we decided to ligate the two inserts first, and then ligate the inserts to the linearized vector.Here we have successfully concatenated the inserts.<br>
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<html><img width = "300" src="https://static.igem.wiki/teams/5291/images/result/electropherogram-of-nadm-nadk-pnta-pntb-pcr-gel.png" /></html><html><img width = "500" src="https://static.igem.wiki/teams/5291/images/parts-lhs/nadm-nadk-pnta-pntb.png" /></html><br>
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<b>Fig.2 Electropherogram of <i>nadM-nadK-pntA-pntB</i> PCR gel.</b>
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Latest revision as of 03:36, 2 October 2024


pBBR1MCS2-nadM-nadK-pntA-pntB

As a plasmid, pBBR1MCS2-nadM-nadK-pntA-pntB can increase the intracellular level of NADPH by introducing four exogenous genes: nadM, nadK, pntA and pntB.

Usage and Biology

To increase the intracellular level of NADPH, we constructed the pBBR1MCS2-nadM-nadK-pntA-pntB plasmid containing four exogenous genes, including nadM, nadK, pntA, and pntB.

Fig.1 The map of plasmid pBBR1MCS2-nadM-nadK-pntA-pntB.

Since the efficiency of homologous recombination of multiple fragments was too low, we decided to ligate the two inserts first, and then ligate the inserts to the linearized vector.Here we have successfully concatenated the inserts.

Fig.2 Electropherogram of nadM-nadK-pntA-pntB PCR gel.


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]