Difference between revisions of "Part:BBa K2505004"

Line 15: Line 15:
 
<!-- -->
 
<!-- -->
  
This gene codes complete iP synthesis enzymes. This gene was introduced to human cell, EA.hy926 cell and produces iP(isopentenyl adenine: kind of cytokinin) as a signaling molecule. <i>atipt4</i> gene and <i>log1</i> gene 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. LOG1 is an enzyme(putative lysine decarboxylase family) that necessary for maturation of iP precursor.
+
<span style="margin-left: 25px;">These two genes(<i>atipt4</i> and <i>log1</i>) are derived from <i>Arabidopais thaliana</i> 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.
  
 
<html>
 
<html>

Revision as of 13:23, 30 October 2017

atipt4-IVS-IRES-log1-polyA

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 1193
    Illegal XhoI site found at 2243
    Illegal XhoI site found at 2255
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 739
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 881


These two genes(atipt4 and log1) 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.

Characterization

===Materials & Methods===