Difference between revisions of "Part:BBa K886001"
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| − | This device was created to express the Cre recombinase enzyme and a target gene under the control of the same promoter (promoter not included in this part). The target gene needs to be amplified by PCR using primers that will insert a loxP and a lox66 sites flanking the sequence in order to integrate it in the lox71 recombination site located upstream the Cre recombinase gene. | + | This device was created to express the Cre recombinase enzyme and a target gene under the control of the same promoter (promoter not included in this part). The target gene needs to be amplified by PCR, using primers that will insert a loxP and a lox66 sites flanking the sequence, in order to integrate it in the lox71 recombination site. For this part the lox71 is located upstream the Cre recombinase gene. An alternative construction is submitted in the part BBa_K886002. |
Once this device is located under the control of the desired promoter, it will express the Cre recombinase that recognizes the lox sites and inserts the target gene under the control of the same promoter. | Once this device is located under the control of the desired promoter, it will express the Cre recombinase that recognizes the lox sites and inserts the target gene under the control of the same promoter. | ||
| − | The Cre expression levels are critical for the recombination, we created two different devices: | + | The Cre expression levels are critical for the recombination, we created two different devices: BBa_K886002 (Cre-Lox71) and BBa_K886001 (Lox71-Cre) for testing if inserting the target gene either upstream or downstream the Cre gene will affect the expression efficiency of the target gene. |
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Revision as of 22:14, 24 September 2012
Recombination Device composed by lox71-Cre recombinase
This device was created to express the Cre recombinase enzyme and a target gene under the control of the same promoter (promoter not included in this part). The target gene needs to be amplified by PCR, using primers that will insert a loxP and a lox66 sites flanking the sequence, in order to integrate it in the lox71 recombination site. For this part the lox71 is located upstream the Cre recombinase gene. An alternative construction is submitted in the part BBa_K886002.
Once this device is located under the control of the desired promoter, it will express the Cre recombinase that recognizes the lox sites and inserts the target gene under the control of the same promoter.
The Cre expression levels are critical for the recombination, we created two different devices: BBa_K886002 (Cre-Lox71) and BBa_K886001 (Lox71-Cre) for testing if inserting the target gene either upstream or downstream the Cre gene will affect the expression efficiency of the target gene.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 396
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 82
- 1000COMPATIBLE WITH RFC[1000]