Rapid Golden Gate assembly of exons from genomic DNA for protein expression in Escherichia coli and Pichia pastoris
Junhao Cheng,
Mingkun Wu,
Ren Zhong,
Dayong Si,
Geng Meng,
Rijun Zhang,
Yueping Zhang
Affiliations
Junhao Cheng
1Laboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science & Technology, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China
Mingkun Wu
1Laboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science & Technology, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China
Ren Zhong
1Laboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science & Technology, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China
Dayong Si
1Laboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science & Technology, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China
Geng Meng
1Laboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science & Technology, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China
Rijun Zhang
1Laboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science & Technology, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China
Yueping Zhang
1Laboratory of Feed Biotechnology, State Key Laboratory of Animal Nutrition, College of Animal Science & Technology, College of Veterinary Medicine, China Agricultural University, Beijing, 100193, China
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.