70550 T7Select®10-3 Cloning Kit

70550
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      Description
      OverviewNovagen's T7Select® Phage Display System is a novel phage display system that takes advantage of the properties of bacteriophage T7. This system is easy to use and has the capacity to display peptides up to about 50 amino acids in size in high copy number (415 per phage), and peptides or proteins up to about 1200 amino acids in low copy number (0.1-1 per phage) or mid-copy number (5-15 per phage). Phage assembly takes place in the E. coli cytoplasm and mature phage are released by cell lysis. Unlike the filamentous systems, peptides or proteins displayed on the surface of T7 do not need to be capable of secretion through the cell membrane, a necessary step in filamentous phage assembly.

      T7 has additional properties that make it an attractive display vector. It is very easy to grow and replicates more rapidly than either bacteriophage l or filamentous phage. Plaques form within 3h at 37°C and cultures lyse 1-2 h after infection, decreasing the time needed to perform the multiple rounds of growth usually required for selection. The T7 phage particle is extremely robust, and is stable to harsh conditions that inactivate other phage. This stability expands the variety of agents that can be used in biopanning selection procedures, which require that the phage remain infective. T7 is an excellent general cloning vector. Purified DNA is easy to obtain in large amounts, a high-efficiency in vitro packaging system has been demonstrated (2), and the DNA is completely sequenced (39,937 bp), so restriction or DNA sequence analysis of clones is quite straightforward.

      The T7Select Phage Display System uses the T7 capsid protein to display peptides or proteins on the surface of the phage. The capsid protein is normally made in two forms, 10A (344 aa) and 10B (397 aa). 10B is produced by a translational frameshift at amino acid 341 of 10A, and normally makes up about 10% of the capsid protein. However, functional capsids can be composed entirely of either 10A or 10B, or of various ratios of the proteins. This finding provided the initial suggestion that the T7 capsid shell could accommodate variation, and that the region of the capsid protein unique to 10B might be on the surface of the phage and could be used for phage display.

      There are 3 basic types of T7Select phage display vectors: the T7Select415 vector for high-copy number display of peptides, the T7Select10 vector for mid-copy number display of peptides or larger proteins, and the T7Select1 vectors for low-copy number display of peptides or larger proteins. In all of the vectors, coding sequences for the peptides or proteins to be displayed are cloned within a series of multiple cloning sites following aa 348 of the 10B protein. The natural translational frameshift site within the capsid gene has been removed, so only a single form of capsid protein is made from these vectors.


      The T7Select System is sold for research use only. Any commercial use of this Product, including drug discovery or development of commercial products, requires a commercial license from EMD Millipore Corporation. Commercial use shall include but not be limited to (1) use of the Product or its components in manufacturing; (2) use of the Product or its components to provide a service, information, or data to others in exchange for consideration; (3) use of the Product or its components (or any derivatives thereof) for therapeutic, diagnostic or prophylactic purposes (including as part of a device, chip, assay or other product); or (4) resale of the Product or its components, whether or not such Product or its components are resold for use in research. Nothing contained herein shall be deemed to represent or warrant that additional third party rights are not required for use of this Product.
      This product contains genetically modified organisms (GMO). Within the EU GMOs are regulated by Directives 2001/18/EC and 2009/41/EC of the European Parliament and of the Council and their national implementation in the member States respectively. This legislation obliges MilliporeSigma to request certain information about you and the establishment where the GMOs are being handled. Click here for Enduser Declaration (EUD) Form.
      Catalogue Number70550
      Brand Family Novagen®
      References
      Product Information
      5 µgT7Select EcoR I/Hind III Vector Arms
      0.2 pmolT7Select Control Insert
      6T7 Packaging Extracts
      1 µgT7Select Packaging Control DNA
      0.2 mlBLT5403, BLT5615, and BL21 glycerol stock
      500 pmolT7SelectUP primer
      500 pmolT7SelectDOWN primer
      Quality LevelMQ100
      Applications
      Biological Information
      Physicochemical Information
      Dimensions
      Materials Information
      Toxicological Information
      Safety Information according to GHS
      Safety Information
      Product Usage Statements
      Storage and Shipping Information
      Ship Code Dry Ice Only
      Toxicity Standard Handling
      Storage ≤ -70°C
      Avoid freeze/thaw Avoid freeze/thaw
      Do not freeze Ok to freeze
      Packaging Information
      Transport Information
      Supplemental Information
      Specifications

      Documentation

      T7Select®10-3 Cloning Kit Certificates of Analysis

      TitleLot Number
      70550

      Citations

      Title
    • Eun-Min Cho, et al. (2007) Phage display cDNA cloning and expression analysis of hydrophobins from the entomopathogenic fungus Beauveria (Cordyceps) bassiana. Microbiology 153, 3438-3447.
    • Lauren R.H. Krumpe, et al. (2006) T7 lytic phage-displayed peptide libraries exhibit less sequence bias than M13 filamentous phage-displayed peptide libraries. Proteomics 6, 4210-4222.
    • James E. Nowak, et al. (2006) Direct production and purification of T7 phage display cloned proteins selected and analyzed on microarrays. Bio/Techniques 40, 220-227.
    • Jan Pilch, et al. (2006) Peptides selected for binding to clotted plasma accumulate in tumor stroma and wounds. Proceedings of the National Academy of Sciences (USA) 103, 2800-2804.
    • Inge Roman, et al. (2006) Hunting interactomes of a membrane protein: obtaining the largest set of VDAC-interacting protein epitopes. Molecular and Cellular Proteomics 5, 1667-1680.
    • Jinguo Chen, et al. (2005) Tachyplesin activates the classic complement pathway to kill tumor cells. Cancer Research 65, 4614-4622.
    • Paul Whitley, et al. (2003) Identification of mammalian Vps24p as an effector of phosphatidylinositol 3,5-bisphosphate-dependent endosome compartmentalization. Journal of Biological Chemistry 278, 38786-38795.
    • Kimmo Porkka, et al. (2002) A fragment of the HMGN2 protein homes to the nuclei of tumor cells and tumor endothelial cells in vivo. Procedings of the National Academy of Science 99, 7444-7449.
    • Paul P. Sche, et al. (1999) Display cloning: functional identification of natural product receptors using cDNA-phage dispaly. Chemistry and Biology 6, 707-716.
    • User Protocols

      Title
      TB178 T7Select® System Manual

      Vector Map

      Title
      TB243 T7Select® 10-3b Vector Map

      Vector Sequence

      Title
      T7Select® 10-3b Vector Sequence

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      Categories

      Life Science Research > Genomic Analysis > Transfection and Protein Expression > Bacterial Expression > Bacterial Expression Vectors
      Life Science Research > Genomic Analysis > DNA Preparation & Cloning > Cloning > Cloning Kits