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Figure 2 | Microbial Cell Factories

Figure 2

From: Strategies for the multiplex mapping of genes to traits

Figure 2

Genome wide fitness measurement techniques. a) Vector library based approaches such as SCALEs [25] start with cloning of random DNA fragments that represent different sites of the genome of interest. This diverse vector library is transformed into the host strain and grown under a defined selective pressure. Following selection the cells are lysed and the vector libraries cut and hybridized to a microarray. The signal intensities from the microarray are then mapped back to the genomic coordinates to identify traits of interest and measure fitness as described in FigureĀ 1. b) Overview of TRMR. An array of rationally designed oligos are cleaved and processed in parallel to produce cassettes with homology arms that guide rational engineering of the entire genome. The TRMR barcodes can then be hybridized to a microarray or PCR amplified and sequenced to determine the fitness of each mutation. This technique provides a versatile method for engineering different functional changes into the genome of interest and rapidly tracking the effects.

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