1. Strain & Vector for use
Wei ZHENG
8th, Jul, 2008

2. Lambda InCh a simple E.Coli plasmid-chromosome shuttle system

3. Advantages No risk of >1 copy of target gene in our machine
Stable integration of a single copy of the plasmid fragment
Most ordinary E.Coli strains and a variety of pBR322-derived Amp-resistant plasmid can be used
λphage as the only specialized vector required

4. General principle
Recombination of the desired plasmid-encoded genes onto lambda InCh
Integration of the recombinant lambda InCh into the chromosome at the lambda attachment site
Deletion of most of the lambda genes, including the att site from the chromosome of the lysogen

6. Two homologous region
“Near ori” – at the region near pBR322 replicative origin
A fragment of the bla gene of pBR322

8. pBR322

9. pUC18

10. How to select the lysogens?
Low frequency transducing (LFT) lysate:
- typically10-3 of the population are recombinant phages, having complete bla gene (Ampr)
Heat-inducible cI857 repressor:
- selection of the temperature- independent derivatives at 42oC

11. E.Coli Strains

12. E.Coli strain BBa_V1001 DH5a Genotype
F-φ80dlacZΔM15 Δ(lacZYA-argF)U169 deoR, recA1 endA1 hsdR17(rk- mk+ phoA supE44 λ- thi-1 gyrA96 relA1

13. Strain: BBa_V1001 DH5a
Vector: -λInCh2
- pUC18

14. More Information Gallery of λInCh Homology Maps
λInCh page
Manual