Rapid profiling of a microbial genome using mixtures of barcoded oligonucleotides View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2010-08

AUTHORS

Joseph R Warner, Philippa J Reeder, Anis Karimpour-Fard, Lauren B A Woodruff, Ryan T Gill

ABSTRACT

A fundamental goal in biotechnology and biology is the development of approaches to better understand the genetic basis of traits. Here we report a versatile method, trackable multiplex recombineering (TRMR), whereby thousands of specific genetic modifications are created and evaluated simultaneously. To demonstrate TRMR, in a single day we modified the expression of >95% of the genes in Escherichia coli by inserting synthetic DNA cassettes and molecular barcodes upstream of each gene. Barcode sequences and microarrays were then used to quantify population dynamics. Within a week we mapped thousands of genes that affect E. coli growth in various media (rich, minimal and cellulosic hydrolysate) and in the presence of several growth inhibitors (beta-glucoside, D-fucose, valine and methylglyoxal). This approach can be applied to a broad range of traits to identify targets for future genome-engineering endeavors. More... »

PAGES

856

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nbt.1653

    DOI

    http://dx.doi.org/10.1038/nbt.1653

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1050549966

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/20639866


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