Difference between revisions of "Part:BBa K1185001:Design"

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===Design Notes===
 
===Design Notes===
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There are no illegal restriction sites in this BioBrick. As the ''hbs'' gene is found naturally in ''B. subtilis'' there is no need for codon optimization. We chose the linker sequence as it is flexible and so does not interfere with the function of the fusion protein.
+
There are no illegal restriction sites in this BioBrick. As the ''hbs'' gene is found naturally in ''B. subtilis'' there is no need for codon optimization. We chose the linker sequence as it is flexible and so does not interfere with the function of the fusion protein. The sfGFP BioBrick has already undergone codon optimization for ''B. subtilis''. We have included a three frame stop codon at the end of our construct.
  
 
===Source===
 
===Source===
  
−
The source of the HBsu coding sequence was the ''B. subtilis'' strain 168 genome, SwissProtTM accession number: P08821. The sfGFP sequence was sourced from BBa_E0040 and the linker from: BBa_K105012.
+
The source of the HBsu coding sequence was the ''B. subtilis'' strain 168 genome, SwissProtTM accession number: P08821. The sfGFP sequence was sourced from BBa_I746916 and the linker from: BBa_K105012.
  
 
===References===
 
===References===
 +
[http://www.ncbi.nlm.nih.gov/pubmed/10224127 Kouji, N., Shou-ichi, Y., Takao, Y. & Kunio , Y., 1999. Bacillus subtilis Histone-like Protein, HBsu, Is an Integral Component of a SRP-like Particle That Can Bind the Alu Domain of Small Cytoplasmic RNA. The Journal of Biological Chemistry , Volume 274, pp. 13569-13576.]
 +
 +
[http://www.ncbi.nlm.nih.gov/pubmed/1902464 Micka, B., Groch, N., Heinemann, U. & Marahiel , M., 1991. Molecular cloning, nucleotide sequence, and characterization of the Bacillus subtilis gene encoding the DNA binding protein HBsu. Journal of Bacteriology, May;173(10), pp. 3191 -3198.]
 +
 +
[http://www.ncbi.nlm.nih.gov/pubmed/1382620 Micka, B. & Marahiel, M., 1992. The DNA-binding protein HBsu is essential for normal growth and development in Bacillus subtilis. Biochimie, 74(7-8), pp. 641-650.]
 +
 +
[http://www.ncbi.nlm.nih.gov/pubmed/10715001 Ross , M. & Setlow, P., 2000. The Bacillus subtilis HBsu protein modifies the effects of alpha/beta-type, small acid-soluble spore proteins on DNA.. Journal of Bacteriology, 182(7), pp. 1942-1948.]

Latest revision as of 16:06, 3 October 2013


HBsu-sfGFP


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 339


Design Notes

There are no illegal restriction sites in this BioBrick. As the hbs gene is found naturally in B. subtilis there is no need for codon optimization. We chose the linker sequence as it is flexible and so does not interfere with the function of the fusion protein. The sfGFP BioBrick has already undergone codon optimization for B. subtilis. We have included a three frame stop codon at the end of our construct.

Source

The source of the HBsu coding sequence was the B. subtilis strain 168 genome, SwissProtTM accession number: P08821. The sfGFP sequence was sourced from BBa_I746916 and the linker from: BBa_K105012.

References

[http://www.ncbi.nlm.nih.gov/pubmed/10224127 Kouji, N., Shou-ichi, Y., Takao, Y. & Kunio , Y., 1999. Bacillus subtilis Histone-like Protein, HBsu, Is an Integral Component of a SRP-like Particle That Can Bind the Alu Domain of Small Cytoplasmic RNA. The Journal of Biological Chemistry , Volume 274, pp. 13569-13576.]

[http://www.ncbi.nlm.nih.gov/pubmed/1902464 Micka, B., Groch, N., Heinemann, U. & Marahiel , M., 1991. Molecular cloning, nucleotide sequence, and characterization of the Bacillus subtilis gene encoding the DNA binding protein HBsu. Journal of Bacteriology, May;173(10), pp. 3191 -3198.]

[http://www.ncbi.nlm.nih.gov/pubmed/1382620 Micka, B. & Marahiel, M., 1992. The DNA-binding protein HBsu is essential for normal growth and development in Bacillus subtilis. Biochimie, 74(7-8), pp. 641-650.]

[http://www.ncbi.nlm.nih.gov/pubmed/10715001 Ross , M. & Setlow, P., 2000. The Bacillus subtilis HBsu protein modifies the effects of alpha/beta-type, small acid-soluble spore proteins on DNA.. Journal of Bacteriology, 182(7), pp. 1942-1948.]