MultiSite Gateway Technology

MultiSite Gateway Pro Plus Kit

What if you could easily and accurately assemble multiple DNA fragments in the order and orientation of you desire? MultiSite Gateway™ Pro technology is the answer, allowing you the flexibility to assemble multiple DNA fragments precisely, efficiently, and directionally in a defined order and orientation and without sub cloning. The full power of Gateway recombinant cloning technology is realized with the MultiSite Gateway Pro technology, which has several protein-expression applications covering the engineering of proteins, pathways, and cells, and provides a highly flexible platform for functional analysis (Table 1).

Table 1. Applications for MultiSite Gateway Technology.

Add promoter/tag element to your Gateway Entry clonesGene knock-down and rescue
Combinationatorial taggingOptimized multi-gene delivery without co-transfection
Construct biological or biochemical pathwaysScreen expression from different promoters
Expression of enzymatic pathwaysShuffle protein coding domains
Expression of multi-subunit protein complexesVariable gene expression levels using different expression elements

Simultaneously assemble 2, 3 or 4 DNA fragments into one vector

Discover how MultiSite Gateway Technology allows you to perform pathway reconstitution, multiple gene expression and regulation, protein interaction studies, and more (Figures 1 and 2). Easily swap and assemble promoters, tags, and genes for your applications, or build libraries of different elements for screening purposes.

  • Simplified cloning of multiple DNA fragments—Gateway recombination eliminates the use of restriction enzymes and ligases
  • Flexible design—clone up to four DNA elements into any Gateway Destination vector
  • Reliable gene delivery—avoid uncertain cotransfection by creating one plasmid containing all your DNA elements

Figure 1. Multiple fragment assembly simplified. Overview the MultiSite Gateway Pro technology.

Figure 2. An example of using MultiSite Gateway Pro Technology to study expression of multiple genes in human cells. Entry clones encoding genes for YFP and CFP and the CMV and EF-1α promoters were recombined into pcDNA™ 6.2/V5-PL-DEST (A, C, and E) or into pcDNA™ 6.2/V5-DEST (B and D). The resulting expression clones were transfected into HeLa cells. Expression was verified under a fluorescence microscope. The plasmid pcDNA™ 6.2/V5-PL-DEST is a promoterless derivative of pcDNA™ 6.2/V5-DEST, which carries the CMV promoter.

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