Difference between revisions of "Part:BBa K343006:Design"

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===Design Notes===
 
===Design Notes===
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1. ''What is the origin of the genetic material used? What does the the genetic materiale do in this origin? Are there uncertainty about the genetical materials function?''
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1. ''Source and function''
  
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The gene was cloned from ''Drosophila melanogaster'' cDNA. The normal function of the gene is to create beta-caroten monooxygenase as outlined above. The function of this gene is well characterized in the literature and there are little reason to suspect it should function otherwise.
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The gene was cloned from ''Drosophila melanogaster'' cDNA aquired from [https://dgrc.cgb.indiana.edu/ Drosophila Genomic Resource Center]. The normal function of the gene is to create beta-caroten 15'15-monooxygenase as outlined above. The function of this gene has been characterized in the literature.
  
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2. ''Modifications before assembly''
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2. ''What modification were done on the genetic materiale before insertion? If anything was modified, what function do you hope to achieve?''
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Appart from addition of BioBrick prefix/suffix no changes were made to the DNA before inserting it into ''E. coli''.
 
Appart from addition of BioBrick prefix/suffix no changes were made to the DNA before inserting it into ''E. coli''.
  
  
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3. ''What vector did you use? Which antibiotic resistance were involved? Which protocol was used to insert the vector?''
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3. ''Vector''
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The gene was inserted into two plasmid backbones, both containing chloramphenicol resistance. Both plasmids are specially made for BioBrick use and as such tested and safe. The plasmid was introduced into ''E. coli'' via chemical transformation.
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4. ''What is the stability of the insert with respect to genetic traits?''
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We have not yet tested the stability of the organism after insertion of our BioBrick.
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5. ''How easily can the insert transfer to other bacteria or lifeforms?''
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We have not tested the vectors ability to transfer the BioBrick to other bacteria.
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The gene was inserted into a pSB1C3 backbone.
  
  
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6. ''Where there safer alternatives to achieve this function? Where there safer alternatives to the host organism and vector used?''
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6. ''Safety considderations''
  
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We considered the gene, the strains of ''E. coli'' and used plasmids as safe. Cell-free systems might have been used, but these have yet to gain the same function as real bacteria.
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We considered the gene, the strains of ''E. coli'' and plasmids used as safe.
  
 
===Source===
 
===Source===

Revision as of 17:26, 26 October 2010

Expresses B-carotene monooxygenase on a constitutive promotor


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 765
    Illegal BamHI site found at 500
    Illegal BamHI site found at 1757
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1361
    Illegal BsaI site found at 1809


Design Notes

1. Source and function

The gene was cloned from Drosophila melanogaster cDNA aquired from Drosophila Genomic Resource Center. The normal function of the gene is to create beta-caroten 15'15-monooxygenase as outlined above. The function of this gene has been characterized in the literature.

2. Modifications before assembly

Appart from addition of BioBrick prefix/suffix no changes were made to the DNA before inserting it into E. coli.


3. Vector

The gene was inserted into a pSB1C3 backbone.


6. Safety considderations

We considered the gene, the strains of E. coli and plasmids used as safe.

Source

The ninaB gene was originally cloned from drosophila melanogaster, and cDNA was aquired for production of the biobrick from the Drosophila Genome Ressource Center. Promotor+rbs brick BBa_J13002 and terminator BBa_B0015 were used.

References