A BioBrick™-Compatible Vector for Allelic Replacement Using the XylE Gene as Selection Marker.

ABSTRACT:

Background

Circular plasmid-mediated homologous recombination is commonly used for marker-less allelic replacement, exploiting the endogenous recombination machinery of the host. Common limitations of existing methods include high false positive rates due to mutations in counter-selection genes, and limited applicability to specific strains or growth media. Finally, solutions compatible with physical standards, such as the BioBrick™, are not currently available, although they proved to be successful in the design of other replicative or integrative plasmids.

Findings

We illustrate pBBknock, a novel BioBrick™-compatible vector for allelic replacement in Escherichia coli. It includes a temperature-sensitive replication origin and enables marker-less genome engineering via two

SUBMITTER: Casanova M 

PROVIDER: S-EPMC4752771 | biostudies-literature | 2016

REPOSITORIES: biostudies-literature