Part:BBa_K1830008
L. lactis Shuttle vector
Lactococcus Lactis shuttle vector part
Usage and Biology
The Lactococcus lactis shuttle vector contains all the components required for transformation, replication and expression of a vector in L. lactis. It only needs to be integrated into the vector that needs to be transformed into the bacteria.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 4023
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Characterization
This part is a necessary requirement for transformation, conservation and replication of a plasmid transformed into L. lactis. We therefore tested by incorporating this part into our main construct. It was then electroporated into L. lactis and selected on chloramphenicol supplemented GM17 media. This test was repeated two times, one with a pSB1C3 plasmid containing only this part, and a second time with a pSB1C3 backbone containing the shuttle vector fused to our main construct. The two transformations were succesfull when plated on selective media. We then decided to check the transformation by extracting our plasmids out of L. lactis. The product was then electrophoresed as well as digested and ran on the same gel, and the functionality of the part was confirmed by the size of the insert and of the entire plasmid. No contamination was detected in the colonies we picked for analysis.
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