EZShuttle™ Recombination Cloning System

 

-Efficient cloning system based on phage lambda recombination, the same principle as Gateway® technology.

Introduction

The GeneCopoeia EZShuttle™ cloning system uses the site-specific recombination machinery between the E. coli and phage lambda genomes for efficient DNA fragment transfer between
plasmids, the same principle as Gateway® technology. The EZShuttle™ system includes the following major components. Click on each tab above for more details:

1. EZRecombinase™ Mixes for catalyzing attL x attR and attB x attP cloning reactions.

2. pEZ™ expression cloning vectors for creating ready-to-use expression clones.

3. EZShuttle™ Gateway® PLUS ORF clones. More than 35,000 human and mouse ORFs in pShuttle™ plasmid backbones.

The EZShuttle™ cloning system is fully compatible with Gateway® cloning, as shown in Table 1.

Table 1. Compatiblilty between components of GeneCopoeia's EZShuttle™ recombination cloning system and Gateway® cloning
technology.

Advantages

  • Complete system of recombination cloning enzymes, expression cloning vectors, and sequence-verified ORF  clones.
  • Highly active for EZRecombinase™ LR Mix and EZRecombinase™ BP Mix.
  • Choice of pEZ™ expression cloning vectors with a wide variety of selection markers and tags for cloning, processing, protein purification and gene function studies.
  • Compatible with existing Gateway® cloning systems.

 

Applications

EZShuttle™ recombination cloning provides a highly fascile system for many powerful applications, including:

  • Create ready-to-express expression clones. Readily shuttle ORFs or other DNA fragments of interest into an expression vector.
  • Protein purification. Express your gene of interest in one or more systems for protein purification, leading to crystallization, antibody generation, DNA binding assays, and
    more.
  • Promoter swapping. Transfer your gene of interest into vectors carrying different promoters.
  • Promoter analysis. Test the function of multiple promoter candidates with a reporter such as luciferase.
  • Gene tagging. Express your gene either untagged or with a multitude of in-frame fusion tags, such as 6XHis, Flag®, GFP, etc. Useful for co-immunoprecipitation,
    subcellular localization, etc.

 

Mechanism of EZShuttle™ cloning

GeneCopoeia's EZShuttle™ recombination-based cloning system for DNA fragment transfer between plasmid vectors is based on the site-specific recombination machinery between the
E. coli and phage lambda genomes, the same principle as Gateway® technology (Figure 1).

 

 

 

 

 

 

Figure 1. The attachment sites and proteins used for integration and excision reactions between E.coli and phage lambda
genomes

 

Note: Gateway® is a registered trademark of Life Technologies.