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Open Access Research article

Molecular characterization of KU70 and KU80 homologues and exploitation of a KU70-deficient mutant for improving gene deletion frequency in Rhodosporidium toruloides

Chong Mei John Koh, Yanbin Liu, Moehninsi, Minge Du and Lianghui Ji*

Author Affiliations

Biomaterials and Biocatalysts Group, Temasek Life Sciences Laboratory, 1 Research Link, National University of Singapore, Singapore 117604, Singapore

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BMC Microbiology 2014, 14:50  doi:10.1186/1471-2180-14-50

Published: 27 February 2014

Additional files

Additional file 1:

Colony colors of ∆car2e after being transformed with a wild type copy of the R. toruloides CAR2 genomic DNA fragment. ∆car2e is a hygromycin sensitive derivative of a CAR2 targeted deletion mutant made by activating the Cre recombinase gene stably integrated into the genome.

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Additional file 2:

Schematic diagram of CAR2 deletion constructs with varied homology length sequence ranging from 50 to 1500 bp used to compare the homologous recombination frequencies between WT and KU70-deficient strain. Restriction enzyme digest sites used for cloning and Southern blot analysis are indicated. The components in the diagram are not drawn to scale.

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Additional file 3:

Comparisons of WT and ∆ku70 strains. (A) Cell morphology; (B) growth rate; (C) sugar consumption rates; (D) fatty acid profiles.

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Additional file 4:

Comparison of gene deletion frequency between different WT and KU70-deficient fungal stains.

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Additional file 5:

(A) Schematic illustration of T-DNA region of pDXP795hptR. Unique restriction enzyme digest sites used are shown. (B) Schematic illustration of CAR2 complementation plasmid within T-DNA region.

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Open Data