Rapid Golden Gate assembly of exons from genomic DNA for protein expression in Escherichia coli and Pichia pastoris

Biotechniques. 2021 Aug;71(2):445-450. doi: 10.2144/btn-2021-0039. Epub 2021 Aug 10.

Abstract

The development of a quick, single-step cloning system for generation of multiexon gene expression constructs is presented. The system allows efficient and cost-effective assembly of multiple exons of interest genes into different expression plasmids in both Escherichia coli and Pichia pastoris. The high cloning efficiency and low cost of the system make it ideal for a novel workflow for the assembly of intron-bearing genes for expression in two different expression hosts.

Keywords: Escherichia coli; Golden Gate; Pichia pastoris; heterologous expression; quick exon assembly.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular
  • DNA
  • Escherichia coli* / genetics
  • Exons
  • Genetic Vectors
  • Genomics
  • Pichia* / genetics
  • Plasmids / genetics
  • Recombinant Proteins / genetics
  • Saccharomycetales
  • Transcriptome*

Substances

  • Recombinant Proteins
  • DNA

Supplementary concepts

  • Komagataella pastoris