Fig. 3: Characterisation of BC11 indicates a genotype-to-phenotype relationship. | Nature Communications

Fig. 3: Characterisation of BC11 indicates a genotype-to-phenotype relationship.

From: Improved betulinic acid biosynthesis using synthetic yeast chromosome recombination and semi-automated rapid LC-MS screening

Fig. 3

a SCRaMbLE strain BC11 exhibited a single recombination event between adjacent LoxPsym sites. This was identified by aligning all raw reads to a pre-SCRaMbLE reference sequence (blue and green lines show individual reads, coloured by direction). The inferred SCRaMbLE event was a deletion of the TIR1 3′ UTR (confirmed by PCR, Supplementary Fig. 6). b Quantitative PCR was performed on the SCRaMbLE BC11 strain in biological and technical triplicates. The mean of three technical repeats for each biological repeat (n = 3) was used to find the biological replicate mean and standard deviation (shown here). c BA titre for yGG066 (negative control, n = 3 biologically independent samples) and BC11 (positive control, n = 3 biologically independent samples) is shown compared to yGG066 d(TIR1-3′) (blue, n = 5 biologically independent samples). Mean and standard deviation shown. d The entire workflow developed in this study is shown. Following mass genome diversification strains are screened using ultra-fast LC-MS to identify the top candidates. These candidates are then subject to downstream characterisation to gain new insight (multiplexed sequencing, genotype-to-phenotype validation, and gene expression analysis). Source data are provided as a Source Data file.

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