Your browser doesn't support javascript.
loading
Multiplexed gene synthesis in emulsions for exploring protein functional landscapes.
Plesa, Calin; Sidore, Angus M; Lubock, Nathan B; Zhang, Di; Kosuri, Sriram.
Afiliación
  • Plesa C; Department of Chemistry and Biochemistry, University of California, Los Angeles (UCLA), Los Angeles, CA, USA.
  • Sidore AM; Department of Chemical and Biomolecular Engineering, UCLA, Los Angeles, CA, USA.
  • Lubock NB; Department of Chemistry and Biochemistry, University of California, Los Angeles (UCLA), Los Angeles, CA, USA.
  • Zhang D; Genomics and Computational Biology Graduate Group, Perelman School of Medicine, University of Pennsylvania, Philadelphia, PA, USA.
  • Kosuri S; Department of Chemistry and Biochemistry, University of California, Los Angeles (UCLA), Los Angeles, CA, USA.
Science ; 359(6373): 343-347, 2018 01 19.
Article en En | MEDLINE | ID: mdl-29301959
Improving our ability to construct and functionally characterize DNA sequences would broadly accelerate progress in biology. Here, we introduce DropSynth, a scalable, low-cost method to build thousands of defined gene-length constructs in a pooled (multiplexed) manner. DropSynth uses a library of barcoded beads that pull down the oligonucleotides necessary for a gene's assembly, which are then processed and assembled in water-in-oil emulsions. We used DropSynth to successfully build more than 7000 synthetic genes that encode phylogenetically diverse homologs of two essential genes in Escherichia coli We tested the ability of phosphopantetheine adenylyltransferase homologs to complement a knockout E. coli strain in multiplex, revealing core functional motifs and reasons underlying homolog incompatibility. DropSynth coupled with multiplexed functional assays allows us to rationally explore sequence-function relationships at an unprecedented scale.
Asunto(s)

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Contexto en salud: 3_ND Problema de salud: 3_neglected_diseases / 3_zoonosis Asunto principal: Proteínas / Genes Sintéticos Idioma: En Revista: Science Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Contexto en salud: 3_ND Problema de salud: 3_neglected_diseases / 3_zoonosis Asunto principal: Proteínas / Genes Sintéticos Idioma: En Revista: Science Año: 2018 Tipo del documento: Article País de afiliación: Estados Unidos
...