Reliable handling of highly A/T-rich genomic DNA for efficient generation of knockin strains of Dictyostelium discoideum.

ABSTRACT:

Background

Social amoeba, Dictyostelium discoideum, is a well-established model organism for studying cellular physiology and developmental pattern formation. Its haploid genome facilitates functional analysis of genes by a single round of mutagenesis including targeted disruption. Although the efficient generation of knockout strains based on an intrinsically high homologous recombination rate has been demonstrated, successful reports for knockin strains have been limited. As social amoeba has an exceptionally high adenine and thymine (A/T)-content, conventional plasmid-based vector construction has been constrained due to deleterious deletion in E. coli.

Results

We describe here a simple and efficient strategy to construct GFP-knockin cassettes by using a linear DNA clonin

SUBMITTER: Mukai A 

PROVIDER: S-EPMC4831088 | biostudies-literature | 2016 Apr

REPOSITORIES: biostudies-literature