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The Invitrogen Gateway Cloning Technology provides a rapid and highly efficient route to protein expression, functional analysis, and cloning/subcloning of DNA segments. Gateway-compatible destination vectors contain the Gateway recombination sites, allowing rapid, efficient transfer of sequences into a variety of expression systems using site-specific recombination.
Gateway cloning technology is especially noted for its utility in protein expression. The flexibility and diverse selection of Destination vectors and host systems is particularly attractive for multidisciplinary protein expression studies.
Host system for protein expression | Gateway Destination vector family |
---|---|
E. coli | • pDEST 14, 15, 17, and 24 • pET160 and pET161 DEST vectors |
Yeast | pYES2-DEST52 |
Insect cells | BaculoDirect C-Term Expression |
Mammalian cells (constitutive expression) | pcDNA mammalian expression vector family |
Mammalian cells (regulated expression) | pT-REx-DEST30, and pT-REx-DEST31 |
Mammalian cells (viral delivery) | ViraPower Lentiviral Expression Systems |
Application | Gateway Destination vector family |
---|---|
Antibody or antigen production | Champion pET Expression systems |
Localization | Vivid Colors pcDNA GFP Destination vector family |
Protein array | Expressway Plus Expression System |
Protein–protein interaction studies | ProQuest Two-Hybrid System using Gateway technology |
Reporter assay | GeneBLAzer pcDNA vector family |
RNAi | GeneBLAzer pcDNA vector family |
High-level inducible mammalian expression
pT-REx-DEST30 andpT-REx-DEST31 are part of the T-REx system, the first inducible mammalian expression system to use the full-length CMV promoter providing high level, regulated expression. There is no toxicity with Thermo Scientific™ T-REx™ because viral transactivators are not used. Easy selection of stable cells using Gibco™ GENETICIN™ is possible due to the presence of the Neomycin resistance gene. Additionally, pT-REx-DEST31 contains a polyhistidine (6xHis) tag allowing for rapid purification on Invitrogen™ ProBond™ resin or detection with an Anti-His Antibody.
Regulated high-level E. coli expression
pET-DEST42 utilizes the T7/lac promoter for regulated, high-level expression in E. coli. The vector has the pBR322 low-copy origin of replication for minimized basal expression level. A C-terminal V5 epitope and polyhistidine (6xHis) tag allow for detection using an Anti-V5 Antibody and an Anti-His Antibody respectively.
To find additional Destination vectors, see our Vector Selection tool