Open AccessRapid Golden Gate assembly of exons from genomic DNA for protein expression in Escherichia coli and Pichia pastoris
Author(s) -
Junhao Cheng,
Mingkun Wu,
Zhong Ren,
Daoyuan Si,
Geng Meng,
Rijun Zhang,
Yueping Zhang
Publication year - 2021
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2021-0039
Subject(s) - pichia pastoris , cloning (programming) , escherichia coli , biology , gene , genomic dna , exon , genetics , plasmid , intron , pichia , computational biology , recombinant dna , microbiology and biotechnology , computer science , programming language
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.