Fig. 2: Engineering and transferring Sc chromosomes into Km. | Nature Communications

Fig. 2: Engineering and transferring Sc chromosomes into Km.

From: Intergeneric chromosomal transfer in yeast results in improved phenotypes and widespread transcriptional responses

Fig. 2: Engineering and transferring Sc chromosomes into Km.

(a) Experimental pipeline. (b) PFGE of chromosomal extracts from Sc-R1, Sc-R3 and the parental Sc strain. (c, d) PCR of markers in the circular chromosomes in KS-R1 (c, represented by one transformant) and KS-R3 (d). Purple box labels the missing marker “7” in KS-R3. See Supplementary Data 1 for marker positions. (e) PFGE of R1 and R3 (arrows) in KS, linearized with NotI and AscI respectively. Ctrl: non-digested KS-R1 chromosomal extracts. λ: Lambda PFG Ladder. (f) Illustration of the 29 kb deletion in KS-R3. Coordinates are based on a manually curated R3 sequence (Supplementary Data 1). (g) Growth curves of KS-R1, KS-R3, and Km-V (Km with an empty vector) in YPD. The values represent mean ± SD (three biological replicates). (h) Spot assay of KS-R1 and KS-R3 in the presence of benomyl (BML), thiabendazole (TBZ), hydroxyurea (HU), methyl methane sulfonate (MMS), camptothecin (CPT) or cycloheximide (CHX). The plates were incubated at 30 degrees for 1 day (1 d) or 3 days (3 d) as indicated. For (b-e), each experiment was repeated three times independently with similar results. Source data are provided as a Source Data file.

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