Difference between revisions of "Part:BBa K733005:Design"

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===Design Notes===
 
===Design Notes===
  
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The mouse BMP-2 DNA sequence has an EcoRI cutting site and this site is within the codon of the DNA sequence. To standardize the biobrick, we try to do a point mutation on mouse BMP-2 after we amplify it from PCR. The purpose is to remove the EcoRI cutting site and remain the correct codon for protein translation of BMP-2.
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The mouse ''Bmp2'' DNA sequence has an EcoRI cutting site and this site is within the codon of the DNA sequence. To standardize the biobrick, we try to do a point mutation on mouse ''Bmp2'' after we amplify it from PCR. This mutation removes the EcoRI cutting site while remaining the correct codon for protein translation of BMP2.
  
 
===Source===
 
===Source===
  
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Extract BMP-2 from mouse genomic DNA by PCR. Extract signal peptide YbdN from ''Bacillus subtilis'' genomic DNA by PCR. Ligation of YbdN and BMP2 achieved by overlapping PCR.
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''Bmp2'' gene is obtained from mouse genomic DNA by PCR. Signal peptide from ''ybdN'' gene is obtained from ''Bacillus subtilis'' genomic DNA by PCR. The recombinant protein YbdN-BMP2 is obtained via overlapping PCR.
  
 
===References===
 
===References===
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Beck, S. E., Jung, B. H., Fiorino, A., Gomez, J., Del Rosario, E., Cabrera, B. L., Huang, S. C., Chow, J. Y. C., & Carethers J.M. (2006). Bone morphogenetic protein signaling and growth suppression in colon cancer. ''The American Journal of Physiology-Gastrointestinal and Liver Physiology'', 291(1), G135-G145.
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Tjalsma, H., Bolhuis, A., Jongbloed, J. D. H., Bron, S., & Dijl, J. M. V. (2000). Signal peptide-dependent protein transport in bacillus subtilis: a genome-based survey of the secretome. ''Microbiology and Molecular Biology Reviews'', 64(3), 515-547.

Latest revision as of 22:56, 26 September 2012

ybdN+Bmp2


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 mouse Bmp2 DNA sequence has an EcoRI cutting site and this site is within the codon of the DNA sequence. To standardize the biobrick, we try to do a point mutation on mouse Bmp2 after we amplify it from PCR. This mutation removes the EcoRI cutting site while remaining the correct codon for protein translation of BMP2.

Source

Bmp2 gene is obtained from mouse genomic DNA by PCR. Signal peptide from ybdN gene is obtained from Bacillus subtilis genomic DNA by PCR. The recombinant protein YbdN-BMP2 is obtained via overlapping PCR.

References

Beck, S. E., Jung, B. H., Fiorino, A., Gomez, J., Del Rosario, E., Cabrera, B. L., Huang, S. C., Chow, J. Y. C., & Carethers J.M. (2006). Bone morphogenetic protein signaling and growth suppression in colon cancer. The American Journal of Physiology-Gastrointestinal and Liver Physiology, 291(1), G135-G145.

Tjalsma, H., Bolhuis, A., Jongbloed, J. D. H., Bron, S., & Dijl, J. M. V. (2000). Signal peptide-dependent protein transport in bacillus subtilis: a genome-based survey of the secretome. Microbiology and Molecular Biology Reviews, 64(3), 515-547.