PUBLICATION
Construction of a series of pCS2+ backbone-based Gateway vectors for overexpressing various tagged proteins in vertebrates
- Authors
- Wang, H.Y., Li, Y., Xue, T., Cheng, N., Du, H.N.
- ID
- ZDB-PUB-161101-7
- Date
- 2016
- Source
- Acta biochimica et biophysica Sinica 48(12): 1128-1134 (Journal)
- Registered Authors
- Li, Yang
- Keywords
- Gateway, pCS2+ vector, pGCS destination vector, tagged proteins, vertebrate organisms
- MeSH Terms
-
- Animals
- Blotting, Western
- HEK293 Cells
- Humans
- Proteins/genetics
- Proteins/metabolism*
- Transfection
- Vertebrates/metabolism*
- PubMed
- 27797719 Full text @ Acta. Biochim. Biophys. Sin (Shanghai)
Citation
Wang, H.Y., Li, Y., Xue, T., Cheng, N., Du, H.N. (2016) Construction of a series of pCS2+ backbone-based Gateway vectors for overexpressing various tagged proteins in vertebrates. Acta biochimica et biophysica Sinica. 48(12):1128-1134.
Abstract
Gateway vectors have been extensively developed to facilitate gene cloning in numerous species; however, a universal system that is compatible for multiple organisms was lacking. As a multipurpose expression vector, pCS2+ backbone-based expression plasmids are widely used for high-level expression of messenger RNAs (mRNAs) or proteins in mammalian/avian culture cells or Xenopus/zebrafish embryos. To date, a suite of vectors with pCS2+ backbone applicable for Gateway cloning system were unavailable yet. Here, we generated a set of Gateway destination vectors, named as pGCS (plasmids of Gateway in pCS2+) vectors, which can be fused to a choice of frequently used amino- or carboxyl-terminal tags, including MYC, HA, FLAG, His, GST, as well as eGFP fluorescent epitope. The systematic generation of this set of pCS2+ backbone-based Gateway destination vectors allows for in vitro recombination of DNA with high speed, accuracy, and reliability compared with the traditional 'digestion-ligation' cloning approach. Thus, our system accelerates the production of functional proteins, which could be widely expressed in a large variety of vertebrate organisms without tediously transferring genes into different expression vectors. Moreover, we make this series of Gateway vectors available to the research community via the non-profit Addgene Plasmid Repository.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping