Part:BBa_K2609026
Lambda Red Recombinases under T7 expression system
This part generates the three Lambda ReD recombinases - exo, gam, beta - when expressed in a T7 expression strain like E. coli BL21(DE3).
Usage and Biology
Biology
The genes exo, gam and beta are a part of Bacteriophage Lambda's genome and is used in recomibination of bacteriophage DNA. The gam protein binds to the RecBCD nuclease of the host, thus protecting linear viral DNA from degradation. The protein exo is a 5' to 3' exonuclease which exposes the ends of linear dsDNA. The protein beta promotes single strand annealing and hence promotes homologous recombination. It is also important in rollin circle DNA replication which comes late in the infective cycle of the lambda phage.
Usage
IISc-Bangalore iGEM 2018
We used the this Lambda RED recombination system with a T4 Endolysin gene (BBa_K2609017) to recombine Bacteriophage T4 and create a lysis deficient phage. This lysis deficient phage triggers its host to produce a monocyte chemoattractant mcp-1 (BBa_K2609000).
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1943
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1460
- 1000COMPATIBLE WITH RFC[1000]
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