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PEL Combinatorial Cloning Platform

The Protein Expression Laboratory Combinatorial Cloning Platform (CCP) is a collection of vectors for use with the Gateway Multisite Recombination system (Life Technologies, Carlsbad, CA) to simplify the construction of expression vectors for wide-ranging areas of genomic and proteomic research.  Some uses of the system are:

- Construction of protein expression constructs with various fusion tags

- Generation of expression constructs with different promoters for in vivo expression

- Production of clones with fluorescent tags for localization experiments

- Generation of constructs for making mutant cell lines or transgenic animals

- Production of vectors for shRNA or miRNA delivery

The advantage of the CCP is in the exquisite specificity of the Multisite Gateway reactions, which permits linkage of multiple elements in a directional fashion, and involves no additional DNA amplification, thus ensuring the accuracy of the DNA sequence of the final products without the need for additional sequencing.  There is also no need for restriction-based cloning processes which have a high rate of failure and may require optimization depending on the sites available in a given clone.

CCP Entry clones have been completely sequence validated and annotated sequence files are provided for all clones.  All clones have also been tested at the PEL to further ensure that they perform as expected.

Please note that the PEL claims no intellectual property rights on these materials—in many cases, the elements in the clones in the collection have IP covered by other companies, and your use of these materials may require a license from the IP holders.  For non-academic users, please be sure to check with the original license holders of clones before using the CCP.

The initial version of the library includes clones for fluorescent and luminescent reporters, epitope and solubility fusion tags, bimolecular fluorescence complimentation (BiFC) fusions, 18 different eukaryotic promoters, and many other useful clones.  In addition, the Destination Vector collection contains two flavors of Gateway destination vectors for E. coli, baculovirus, mammalian, and lentiviral expression, along with materials for generating your own CCP-style destination vectors.

Academic and government researchers can obtain the materials via a simple Materials Transfer Agreement (MTA).  Industry researchers can license the technology from the NIH Office of Technology Transfer by contacting Sury Vepa at vepas@mail.nih.gov

Files:

PEL-CCP Overview (PDF)

PEL-CCP Multisite Protocol (PDF)

PEL-CCP Clone Library 1 (PDF)

PEL-CCP Destination Vectors (PDF)

Page last updated January 7, 2013 @ 2:44 pm