Part:BBa_K343006:Design
Expresses B-carotene monooxygenase on a constitutive promotor
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 765
Illegal BamHI site found at 500
Illegal BamHI site found at 1757 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1361
Illegal BsaI site found at 1809
Design Notes
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?
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.
2. What modification were done on the genetic materiale before insertion? If anything was modified, what function do you hope to achieve?
Appart from addition of BioBrick prefix/suffix no changes were made to the DNA before inserting it into E. coli.
3. What vector did you use? Which antibiotic resistance were involved? Which protocol was used to insert the vector?
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.
4. What is the stability of the insert with respect to genetic traits?
We have not yet tested the stability of the organism after insertion of our BioBrick.
5. How easily can the insert transfer to other bacteria or lifeforms?
We have not tested the vectors ability to transfer the BioBrick to other bacteria.
6. Where there safer alternatives to achieve this function? Where there safer alternatives to the host organism and vector used?
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.
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.