pLDR8 [WM2269] (ATCC® 77357)

Organism: bacteriophage lambda  /  Clone Type: Clone  /  Depositors: W Messer

Permits and Restrictions

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Designations pLDR8 [WM2269]
Species bacteriophage lambda
Depositors W Messer
Applications
In another host, produces protein integrase
In another host, produces protein repressor protein cI857 mutation
Vector
Construct size (kb): 7.6
Insert
DNA: genomic
Gene product: integrase [int]
Target Gene: integrase, repressor protein cI857 mutation
Media Medium 1236: LB Medium (ATCC medium 1065) with 25 mcg/ml kanamycin
Related Products
Component of: ATCC 77371
Biosafety Level 1
Comments
Restriction digests of the clone give the following sizes (kb): EcoRI/PstI--5.4, 2.2; BamHI--4.2, 3.4; PstI--7.6; HindIII--5.7, 1.9.
Temperature-resistant derivatives arise frequently. It is important to isolate a clone that is temperature-sensitive and kanamycin resistant before use.
pLDR8 is compatible with, and provides Int function for, the vectors pLDR9 (ATCC 77358), pLDR10 (ATCC 77359) and pLDR11 (ATCC 77360).
Expression of int in pLDR8 is regulated by lambda PR, by the temperature sensitive cI857 repressor. Replication of pLDR8 is also temperature sensitive.
Therefore, at 30C replication of the plasmid is normal and int is not expressed. At 42C, int is expressed but pLDR8 is no longer replicated, resulting in loss of int after a few cell generations.
The order of the major features in this plasmid is: EcoRI - pSC101 ori - neoR - PstI - cI857 - PR promoter - int.
References

Diederich L, et al. New cloning vectors for integration in the lambda attachment site attB of the Escherichia coli chromosome. Plasmid 28: 14-24, 1992. PubMed: 1387714

Walter Messer, personal communication

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