Part:BBa_K2325104
N20-sgRNA function device in pTarget
The BBa_K2325104 is a construct contains N20 sequence selected from the genome of T7 phage, with a constitutive promoter, which is constructed in the CRISPR/Cas system to make the E.coli be able to confer resistance to T7 phage.
Usage and Biology
The pTarget-N20 processing this device is co-transfected with pCas into E.coli BL21(DE3) strain, resulting in BL21(DE3)(pCas+pTarget-N20).
Gel analysis
Figure 1. Growth curve of BL21(DE3) (pCas+pTarget) with and without the T7 phage infection and E.coli BL21(DE3)(negative control) with the T7 phage infection in LB media.
The results convinced that the recombinant strain functioned quite well to resist the phage in LB, and the efficiency of resistance reached nearly 100%.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 7
Illegal NheI site found at 30 - 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 41
| None |