Source:
Generated By: https://synbiohub.org/public/igem/igem2sbol/1
Created by: HO Yuan Heng Trevor
Date created: 2012-08-25 11:00:00
Date modified: 2015-05-08 01:12:34
pTE
| Types | DnaRegion |
| Roles | plasmid Plasmid |
| Sequences | BBa_K524204_sequence (Version 1) |
Description
pTE is a template plasmid for generating linear DNA substrate for E. coli homologous recombination through lambda RED recombination system. The plasmid only contain the kanamycin resistant gene and the R6K origin of replication.In using lambda RED recombination, a linear piece of dsDNA is necessary and serves as the substrate for homologous recombination. Such linear dsDNA (in which flanking regions of homologies are incorporated by extending primers used) is usually amplified from a plasmid with a resistance gene as the marker. During purification of the amplicon, plasmids serving as templates can easily contaminate the linear dsDNA, and they would give false positives during isolation of recombinants. (Datsenko and Wanner) To avoid this problem, Datsenko and Wanner introduced a plasmid pKD3 with R6K origin of replication that only allows propagation in permissive host with pi protein provided in trans. Such template plasmids will not propagate in the designated hosts for recombination.
The pTE plasmid is a direct mimicking of the pKD4 plamsmid by using available parts from the Registry, for the purpose of generating linear dsDNA that contamination by template plasmid would not cause a problem
The necessity for using pTE to amplify linear DNA substrate lies in the fact that template plasmids