Difference between revisions of "Part:BBa K1189009:Design"
 
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Note: The Bsa1 cut site has NOT been removed. This part has been designed in the RFC 25 Freiburg fusion backbone and can be used to make fusion proteins. The ATG codon is incorporated in the backbone before the NgoMIV restriction cut site.  
 
Note: The Bsa1 cut site has NOT been removed. This part has been designed in the RFC 25 Freiburg fusion backbone and can be used to make fusion proteins. The ATG codon is incorporated in the backbone before the NgoMIV restriction cut site.  
  
−
========
+
======
 
TCT: Original serine - mutated to TCC: serine     
 
TCT: Original serine - mutated to TCC: serine     
−
========
+
======
 
Mutagenesis Primers
 
Mutagenesis Primers
−
========
+
======
 
F: 5’GGAGCCGGTGAGCGTGGGTCCCGCGGTATCATTGCAGC 3’
 
F: 5’GGAGCCGGTGAGCGTGGGTCCCGCGGTATCATTGCAGC 3’
−
========
+
======
 
R: 5’GCTGCAATGATACCGCGGGACCCACGCTCACCGGCTCC 3’
 
R: 5’GCTGCAATGATACCGCGGGACCCACGCTCACCGGCTCC 3’
 +
 +
===References===
 +
<li>Kong, Y., Yao, H., Ren, H., Subbian, S., Cirillo, S. L. G., Sacchettini, J. C., … Cirillo, J. D. (2010). Imaging tuberculosis with endogenous beta-lactamase reporter enzyme fluorescence in live mice. <i>Proceedings of the National Academy of Sciences of the United States of America, 107</i>(27), 12239–44. doi:10.1073/pnas.1000643107
 +
<li>Moore, J. T., Davis, S. T., & Dev, I. K. (1997). The development of beta-lactamase as a highly versatile genetic reporter for eukaryotic cells. <i>Analytical Biochemistry, 247</i>(2), 203–9. doi:10.1006/abio.1997.2092
 +
<li>Qureshi, S. (2007). β-Lactamase: an ideal reporter system for monitoring gene expression in live eukaryotic cells. <i>BioTechniques, 42</i>(1), 91–96. doi:10.2144/000112292

Latest revision as of 00:38, 26 October 2013

Beta-lactamase with His Tag


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

Note: The Bsa1 cut site has NOT been removed. This part has been designed in the RFC 25 Freiburg fusion backbone and can be used to make fusion proteins. The ATG codon is incorporated in the backbone before the NgoMIV restriction cut site.

==

TCT: Original serine - mutated to TCC: serine

==

Mutagenesis Primers

==

F: 5’GGAGCCGGTGAGCGTGGGTCCCGCGGTATCATTGCAGC 3’

==

R: 5’GCTGCAATGATACCGCGGGACCCACGCTCACCGGCTCC 3’

References

  • Kong, Y., Yao, H., Ren, H., Subbian, S., Cirillo, S. L. G., Sacchettini, J. C., … Cirillo, J. D. (2010). Imaging tuberculosis with endogenous beta-lactamase reporter enzyme fluorescence in live mice. Proceedings of the National Academy of Sciences of the United States of America, 107(27), 12239–44. doi:10.1073/pnas.1000643107
  • Moore, J. T., Davis, S. T., & Dev, I. K. (1997). The development of beta-lactamase as a highly versatile genetic reporter for eukaryotic cells. Analytical Biochemistry, 247(2), 203–9. doi:10.1006/abio.1997.2092
  • Qureshi, S. (2007). β-Lactamase: an ideal reporter system for monitoring gene expression in live eukaryotic cells. BioTechniques, 42(1), 91–96. doi:10.2144/000112292