Difference between revisions of "Part:BBa K886003"
| Line 6: | Line 6: | ||
However, in order to have this recombination a promoter is needed (there isn't a specific promoter for this purpose). It will be responsable for the Cre recombinase and the target gene expression. | However, in order to have this recombination a promoter is needed (there isn't a specific promoter for this purpose). It will be responsable for the Cre recombinase and the target gene expression. | ||
| − | Knowing that Cre expression levels are critical for the recombination, we've created two different devices: | + | Knowing that Cre expression levels are critical for the recombination, we've created two different devices: BBa_K886001 (Lox71-Cre) and BBa_K886003 (Cre-Lox71) for testing if the insertion of the target gene either upstream or downstream the Cre gene will affect its expression efficiency. |
As described above, in this part the lox71 is located downstream the Cre recombinase gene. An alternative construction can be found in the part BBa_K886001. | As described above, in this part the lox71 is located downstream the Cre recombinase gene. An alternative construction can be found in the part BBa_K886001. | ||
Revision as of 02:35, 25 September 2012
Recombination Device composed by Cre-lox71
This device was created to express the Cre recombinase enzyme and a target gene, both under the control of the same promoter (promoter not included in this part). The target gene needs to be amplified by PCR with primers capable of inserting a loxP and a lox66 sites on both sides of the PCR product, "floxing" (flanking) it. This construction allows the target gene to be inserted into the lox71 recombination site of the device. The process occurs via Cre recombinase recognition and interaction with the lox sites.
However, in order to have this recombination a promoter is needed (there isn't a specific promoter for this purpose). It will be responsable for the Cre recombinase and the target gene expression.
Knowing that Cre expression levels are critical for the recombination, we've created two different devices: BBa_K886001 (Lox71-Cre) and BBa_K886003 (Cre-Lox71) for testing if the insertion of the target gene either upstream or downstream the Cre gene will affect its expression efficiency.
As described above, in this part the lox71 is located downstream the Cre recombinase gene. An alternative construction can be found in the part BBa_K886001.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 356
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 42
- 1000COMPATIBLE WITH RFC[1000]