Difference between revisions of "Part:BBa K2505005"
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This part produces complete iP(isopentenyl adenine: kind of cytokinin) synthesis enzymes induced by chimeric transactivater. This part was introduced human cell('''EA.hy926''') then the cells produced iP when receiving signaling molecule, C8(AHL: quorum sensing signal) from E. coli. The genes(<i>log1</i> and <i>atipt4</i>) is derived from <i>A.thaliana</i>, optimized for <i>H.sapiens</i> codon in reference to the codon usage. AtIPT4 is an enzyme(adenylate dimethylallyltransferase '''[EC:2.5.1.112]''': cytokinin synthase) that necessary for a synthesis of iP(isopentenyl adenine: kind of cytokinin) precursor. LOG1 is an enzyme(putative lysine decarboxylase family) that necessary for a maturation of iP precursor. These enzymes are induced by TraR-C8 complex binding to tra-Box(TraR binding site:Part:BBa_K553021)
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These two genes are derived from Arabidopais thaliana and encode enzymes  necessary for synthesizing iP (isopentenyladenine) in mammalian cells. 
 +
iP is a kind of cytokinins that are signaling molecules (Phytohormones)
 +
in plants and play important roles in cell growth and differentiation.
 +
When these genes are introduced to human cells, EA.hy926, the cells
 +
produce iP heterologously. The DNA sequences of these genes are 
 +
optimized for expressing in human cells considering the codon usage.
 +
AtIPT4 has the adenylate dimethylallyltransferase (EC:2.5.1.112:
 +
cytokinin synthase) activity and catalyzes the transfer of an 
 +
isopentenyl group from dimethylallyl diphosphate (DMAPP) to ATP and ADP, 
 +
producing cytokinin nucleotides. Note that cytokinin nucleotides are the 
 +
immature form. LOG1 has the phosphoribohydrolase activity and converts 
 +
inactive cytokinin nucleotides to the biologically active free-base forms.
 
  CMVmin (minimal CMV promoter: BBa_553022).
 
  CMVmin (minimal CMV promoter: BBa_553022).
  

Revision as of 00:12, 31 October 2017

(tra box)7-CMVmin-atipt4-IVS-IRES-log1-polyA

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1442
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 26
    Illegal BglII site found at 72
    Illegal BglII site found at 118
    Illegal BglII site found at 164
    Illegal BglII site found at 210
    Illegal BglII site found at 256
    Illegal BglII site found at 302
    Illegal BamHI site found at 476
    Illegal XhoI site found at 2728
    Illegal XhoI site found at 2740
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1220
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1362


These two genes are derived from Arabidopais thaliana and encode enzymes necessary for synthesizing iP (isopentenyladenine) in mammalian cells. iP is a kind of cytokinins that are signaling molecules (Phytohormones) in plants and play important roles in cell growth and differentiation. When these genes are introduced to human cells, EA.hy926, the cells produce iP heterologously. The DNA sequences of these genes are optimized for expressing in human cells considering the codon usage. AtIPT4 has the adenylate dimethylallyltransferase (EC:2.5.1.112: cytokinin synthase) activity and catalyzes the transfer of an isopentenyl group from dimethylallyl diphosphate (DMAPP) to ATP and ADP, producing cytokinin nucleotides. Note that cytokinin nucleotides are the immature form. LOG1 has the phosphoribohydrolase activity and converts inactive cytokinin nucleotides to the biologically active free-base forms. 

CMVmin (minimal CMV promoter: BBa_553022).

Characterization

Materials & Methods