Part:BBa_K4019005
pMG36e
The vector is based on pWVO1, originally obtained from L. lactis subsp. cremoris Wg2, which also replicates in E. coli and B. siitilis. The part of the vector containing the pWVO1 origin of replication was obtained from pGK11. This plasmid was reported by Van De Guchte, M. et.al.
Usage and Biology
To confirm that our backbone plasmid works in L.lactis, we measured transformation efficiency. Lactococcus lactis subsp. cremoris NZ9000 was harvested in MRS medium and collected by centrifugation. Collected cells are washed with HEPES,sucrose buffer and suspended. The pMG36e was transformed into L.lactis by electroporation at 1500V/2000V/2500V. Transformants are spread on MRS agar plates with and without 4 µg/mL erythromycin. Colonies could be seen on the plates of 1500 V. Colony forming units was calculated as follows, and it was 1000 CFU/ug. No colonies were observed at 2000 V and 2500 V, but colonies gradually appeared over 2-6 days at 1500 V.
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 2476
Illegal XbaI site found at 2709
Illegal PstI site found at 2721 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 2476
Illegal NheI site found at 1263
Illegal PstI site found at 2721 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 2476
- 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 2476
Illegal XbaI site found at 2709
Illegal PstI site found at 2721 - 25INCOMPATIBLE WITH RFC[25]Illegal EcoRI site found at 2476
Illegal XbaI site found at 2709
Illegal PstI site found at 2721
Illegal AgeI site found at 2809
Illegal AgeI site found at 2895 - 1000COMPATIBLE WITH RFC[1000]
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