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pET-31b(+) DNA, AlwN I Digest
This kit is available as a convenient supplement to the pET Peptide Expression System 31b, which is designed for expression and purification of target peptides (1). In pET-31b(+), peptide coding sequences are placed downstream of a 125 amino acid (aa) ketosteroid isomerase (KSI) gene and upstream of a His•Tag® sequence. Use of the unique AlwN I cloning site enables unidirectional insertion of coding sequences immediately adjacent to a methionine residue. For high-yield production, small (10–25 aa) peptide coding sequences can be cloned as tandem repeats interspersed with single methionine residues. The KSI-peptide-His•Tag fusion protein is expressed at high levels in the presence of T7 RNA polymerase provided by the recA–, protease deficient BLR(DE3)pLysS host. The fusion protein is purified in one step under denaturing conditions using one of Novagen's His•Bind® metal chelation resins. It is then cleaved with CNBr to release the His•Tag peptide, insoluble KSI, and monomer peptide units each terminating with a homoserine lactone. The purified peptide can be easily converted to an amide or homoserine, or reacted with other compounds, such as analogs of fluorescein and biotin, to produce C-terminal derivatized peptides. The system has been used to make biologically active peptides of 14–67 aa at yields of > 50 mg purified peptide per liter of culture (1).
The AlwN I digested, dephosphorylated vector is ready to accept appropriate inserts containing ...ATG-3′ and 3′-TAC... overhangs on the top and bottom strands, respectively. A Control Insert is provided to verify performance and ensure optimal cloning efficiency.
Features
• High yield expression of peptides and small proteins
• His•Tag sequence facilitates protein purification
• Prepared vector facilitates cloning with minimal background
• Special host strain inhibits degradation of expressed proteins..
The pET System allows for expression of target genes under control of the T7 promoter and is covered by US Patent 5,693,489. This patent is licensed by EMD Millipore Corporation from Brookhaven National Labs. Commercial entities need to obtain a license from Brookhaven National Labs prior to use of this Product. Ccommercial use of this Product also 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.
产品信息
Avoid freeze/thaw 是的
Components
• 2 µg AlwN I digested, dephosphorylated pET-31b(+) DNA
• 2 pmol AlwN I Control Insert
• 0.2 ml Induction Control H glycerol stock
商店和运输信息
Storage ≤ -70°C
Ship Shipped with Blue Ice or with Dry Ice
Standard Handling |