Rapid mapping of insertional mutations to probe cell wall regulation in Cryptococcus neoformans.

ABSTRACT: Random insertional mutagenesis screens are important tools in microbial genetics studies. Investigators in fungal systems have used the plant pathogen Agrobacterium tumefaciens to create tagged, random mutations for genetic screens in their fungal species of interest through a unique process of trans-kingdom cellular transconjugation. However, identifying the locations of insertion has traditionally required tedious PCR-based methods, limiting the effective throughput of this system. We have developed an efficient genomic sequencing and analysis method (AIM-Seq) to facilitate identification of randomly generated genomic insertions in microorganisms. AIM-Seq combines batch sampling, whole genome sequencing, and a novel bioinformatics pipeline, AIM-HII, to rapidly identify sites of genomic i

SUBMITTER: Esher SK 

PROVIDER: S-EPMC4693612 | biostudies-literature | 2015 Sep

REPOSITORIES: biostudies-literature