Difference between revisions of "Part:BBa K1352010:Experience"

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(→‎Applications of BBa_K1352010)
 
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Figure 1. Physiology of BBa_K1352000 in response to 0.2% arabinose induction.
 
Figure 1. Physiology of BBa_K1352000 in response to 0.2% arabinose induction.
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Left: BioBrick BBa_K1352000; right: negative control.  
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Left: BioBrick BBa_K1352000; right: negative control (untransformed ''E.coli'').  
  
 
'''Conclusion:'''
 
'''Conclusion:'''
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BBa_K1352000 is the modularized version of BBa_K759001, which is compatible with iGEM Assemly Standard RCF10. Removal of the 6 additional PstI sites through Site-Directed Mutagenesis was confirmed by gel electrophoresis and sequence analysis, with no indels being introduced within the coding region of the construct.  
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BBa_K1352010 is the correct, iGEM RFC10-compatible version of BBa_K759001. In our experience, K759001 is not RFC10 compatible, having 6 Pst1 sites (see 'Design' for K1352010). During the construction of K1352010, the removal of the 6 additional PstI sites through Site-Directed Mutagenesis was confirmed by gel electrophoresis and sequence analysis, with no indels being introduced within the coding region of the construct.
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Antigen 43 protein expression of biobrick BBa_K1352000 was shown not to be affected by the     PCR-based Site-Directed Mutagenesis process. The biobrick works biologically as intended after modification and has the same response to arabinose induction: formation of culture aggregation.
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Antigen 43 protein expression of biobrick BBa_K1352010 was shown not to be affected by the PCR-based Site-Directed Mutagenesis process. The biobrick works biologically as intended after modification and has the same response to arabinose induction: formation of culture aggregation.
  
 
===User Reviews===
 
===User Reviews===

Latest revision as of 11:04, 2 October 2014


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1352010

When this Ag43 basic part is expressed under control of the arabinose responsive promoter in composite part K1352000, the E.coli culture aggregates (as shown below, Figure 1), demonstrating the engineered gene now lacking PstI sites is functional.

K1352000 aggregation.jpg

Figure 1. Physiology of BBa_K1352000 in response to 0.2% arabinose induction. Left: BioBrick BBa_K1352000; right: negative control (untransformed E.coli).

Conclusion: BBa_K1352010 is the correct, iGEM RFC10-compatible version of BBa_K759001. In our experience, K759001 is not RFC10 compatible, having 6 Pst1 sites (see 'Design' for K1352010). During the construction of K1352010, the removal of the 6 additional PstI sites through Site-Directed Mutagenesis was confirmed by gel electrophoresis and sequence analysis, with no indels being introduced within the coding region of the construct.

Antigen 43 protein expression of biobrick BBa_K1352010 was shown not to be affected by the PCR-based Site-Directed Mutagenesis process. The biobrick works biologically as intended after modification and has the same response to arabinose induction: formation of culture aggregation.

User Reviews

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