Difference between revisions of "Part:BBa K404106:Design"

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===Design Notes===
 
===Design Notes===
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The phTERT was created by PCR amplifying the phTERT with overhanging RFC10 iGEM restriction sites and ligated it into pSB1C3. The pHTERT does not contain any iGEM restriction sites being compatible with the RFC25 and RFC10 standard.
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===Source===
 
===Source===
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<p style="text-align:justify;">
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At the 3´end of the human chromosomes the DNA sequence” TTAGGG” can be found in a repetitively manner. The DNA polymerase cannot elongate at this overhanging regions because the overhangs do not provide any template strand for DNA polymerases. In most somatic cells, except for germ cells, the telomerase activity is shut off (Cech 2000).
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Human telomerase is a reverse transcriptase which synthesizes telomeric DNA sequences. The ribonucleoprotein consists of two subunits – the catalytic protein subunit human telomerase reverse transcriptase which catalyzes the polymerization of the nucleotides onto the chromosomes and the RNA component which serve as the template for the synthesis of the telomeric repeat sequences (Wick et al. 1999). Human telomerase reverse transcriptase is driven by the phTERT promoter.
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===References===
 
===References===
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<p style="text-align:justify;">
 
<b>Cech, T.</b>, 2000. Life at the End of the Chromosome: Telomeres and Telomerase. Angewandte Chemie (International ed. in English), 39(1), 34-43. Available at: http://www.ncbi.nlm.nih.gov/pubmed/10649348.<br />
 
<b>Cech, T.</b>, 2000. Life at the End of the Chromosome: Telomeres and Telomerase. Angewandte Chemie (International ed. in English), 39(1), 34-43. Available at: http://www.ncbi.nlm.nih.gov/pubmed/10649348.<br />
 
<b>Kyo, S. et al.</b>, 2008. Understanding and exploiting hTERT promoter regulation for diagnosis and treatment of human cancers. Cancer science, 99(8), 1528-38. Available at: http://www.ncbi.nlm.nih.gov/pubmed/18754863.<br />
 
<b>Kyo, S. et al.</b>, 2008. Understanding and exploiting hTERT promoter regulation for diagnosis and treatment of human cancers. Cancer science, 99(8), 1528-38. Available at: http://www.ncbi.nlm.nih.gov/pubmed/18754863.<br />
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<b>Takakura, M. et al.</b>, 2005. Function of AP-1 in Transcription of the Telomerase Reverse Transcriptase Gene ( TERT ) in Human and Mouse Cells. Society, 25(18), 8037-8043.<br />
 
<b>Takakura, M. et al.</b>, 2005. Function of AP-1 in Transcription of the Telomerase Reverse Transcriptase Gene ( TERT ) in Human and Mouse Cells. Society, 25(18), 8037-8043.<br />
 
<b>Wick, M., Zubov, D. & Hagen, G.</b>, 1999. Genomic organization and promoter characterization of the gene encoding the human telomerase reverse transcriptase (hTERT). Gene, 232(1), 97-106. Available at: http://www.ncbi.nlm.nih.gov/pubmed/10333526.
 
<b>Wick, M., Zubov, D. & Hagen, G.</b>, 1999. Genomic organization and promoter characterization of the gene encoding the human telomerase reverse transcriptase (hTERT). Gene, 232(1), 97-106. Available at: http://www.ncbi.nlm.nih.gov/pubmed/10333526.
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</p>

Latest revision as of 23:31, 22 October 2010

phTERT promoter


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]


Design Notes

The phTERT was created by PCR amplifying the phTERT with overhanging RFC10 iGEM restriction sites and ligated it into pSB1C3. The pHTERT does not contain any iGEM restriction sites being compatible with the RFC25 and RFC10 standard.

Source

At the 3´end of the human chromosomes the DNA sequence” TTAGGG” can be found in a repetitively manner. The DNA polymerase cannot elongate at this overhanging regions because the overhangs do not provide any template strand for DNA polymerases. In most somatic cells, except for germ cells, the telomerase activity is shut off (Cech 2000). Human telomerase is a reverse transcriptase which synthesizes telomeric DNA sequences. The ribonucleoprotein consists of two subunits – the catalytic protein subunit human telomerase reverse transcriptase which catalyzes the polymerization of the nucleotides onto the chromosomes and the RNA component which serve as the template for the synthesis of the telomeric repeat sequences (Wick et al. 1999). Human telomerase reverse transcriptase is driven by the phTERT promoter.

References

Cech, T., 2000. Life at the End of the Chromosome: Telomeres and Telomerase. Angewandte Chemie (International ed. in English), 39(1), 34-43. Available at: http://www.ncbi.nlm.nih.gov/pubmed/10649348.
Kyo, S. et al., 2008. Understanding and exploiting hTERT promoter regulation for diagnosis and treatment of human cancers. Cancer science, 99(8), 1528-38. Available at: http://www.ncbi.nlm.nih.gov/pubmed/18754863.
Maida, Y. et al., 2002. Direct activation of telomerase by EGF through Ets-mediated transactivation of TERT via MAP kinase signaling pathway. Oncogene, 21(26), 4071-9. Available at: http://www.ncbi.nlm.nih.gov/pubmed/12037663.
Nishi, H. et al., 2004. Hypoxia-inducible factor 1 mediates upregulation of telomerase (hTERT). Molecular and cellular biology, 24(13), 6076-83.
Takakura, M. et al., 2005. Function of AP-1 in Transcription of the Telomerase Reverse Transcriptase Gene ( TERT ) in Human and Mouse Cells. Society, 25(18), 8037-8043.
Wick, M., Zubov, D. & Hagen, G., 1999. Genomic organization and promoter characterization of the gene encoding the human telomerase reverse transcriptase (hTERT). Gene, 232(1), 97-106. Available at: http://www.ncbi.nlm.nih.gov/pubmed/10333526.