Product Information

The BIP-300 plasmid allows you to compare your pAN or pAC AviTagTM fusion gene"s protein expression against a confirmed AviTag^TM fusion product under the same growth and induction conditions. When transformed into the same E. coli host strain as your own pAN or pAC AviTagTM fusion plasmid construct, and induced in parallel, the BIP-300 Positive Control Plasmid will provide a measure by which to compare your transformation efficiency, growth and antibiotic maintenance conditions, and induction of your plasmid"s gene-fusion construct. The BIP-300 construct consists of a modified β-galactosidase gene (8 amino acids were deleted from its N-terminus) cloned into Avidity"s pAN5 AviTag^TM fusion vector at the Multiple Cloning Site. The resulting AviTag^TM-protein fusion (AviTag-β-gal), as are all the pAN and pAC fusion products, is under the tight control of the Trc promoter system and expressed upon IPTG induction. The plasmid is maintained with ampicillin. An example induction protocol is given under Protocols. This protocol uses the Avidity AVB101 E. coli strain containing the BirA over-expressing pACYC-184 plasmid construct (pBirAcm) as the host strain. It is initially requires chloramphenicol for maintenance in addition to the ampicillin required to maintain the pAN or pAC plasmid construct. If you are using a different host strain please adjust the protocol accordingly.

Supplied as 1 vial of Positive Control Plasmid DNA; 2µg at 1mg/mL in nuclease-free water (2µL). Store at -20°C.    Avidity带有AviTag标签的人类全长ORF克隆已经被构建在多种T7和CMV启动子表达系统,可以立即购买使用。AviTag标签蛋白可以被体内或体外的生物素连接酶生物素化，而且抗生物素蛋白或链霉亲和素可以专一地与生物素结合，正基于这两个反应，AviTag?技术可以应用于蛋白的固定，纯化和显影。 虽然Avidity是一个小公司，科学带来的好处及其AviTag已得到认可。目前， AviTag技术已经在22个国家的世界十大制药公司和研究人员中得到认可。  Avidity Avitag™ Technology Avidity develops and sells molecular affinity tools for connecting molecules. Our patented AviTag™ technology employs a highly targeted enzymatic conjugation of a single biotin on a unique 15 amino acid peptide tag using the biotin ligase (BirA) from E. coli. AviTag™ sequence is GLNDIFEAQKIEWHE. Oriented on streptavidin-coated surfaces, this creates an ideal presentation for molecular binding interactions. The scientific benefits of Avidity's AviTag Technology have garnered notice. Currently, AviTag technology is licensed by seven of the world's top ten pharmaceutical companies and is used by researchers in 22 countries. Company History Avidity, LLC was founded in 1996 by Millard Cull, Larry Lansing, Dr. Ron Gill, and Dr. Mark Seville to develop peptide tags and molecular biology products based upon the extraordinary affinity of biotin for avidin or streptavidin. While working at Affymax as a researcher, Millard was involved in the discovery of the AviTag technology, saw its commercial potential, and obtained the exclusive rights from Affymax. He started Avidity as a reagent company that sublicenses the right to use the AviTag technology to other companies and sells products for use with the technology. AviTag History AviTag is a patented biotin-tagging technology discovered at Affymax by Dr. Peter Schatz from a Directed Evolution approach called "Peptides-on-Plasmids." The Directed Evolution approach identified a number of peptide sequences that can be biotinylated by the BirA enzyme of E. coli, and the best of these sequences was called the AviTag. E. coli enzyme BirA biotinylates the 15 amino acid AviTag twice as fast as its natural substrate, the Biotin Carboxyl Carrier Protein (BCCP). The enzymatic biotinylation of proteins using AviTag technology is preferred over chemical biotinylation of proteins because BirA enzyme gently, and highly specifically, labels the lysine residue of the AviTag. Prior to the AviTag, the smallest biotin-accepting domain was comprised of 75 amino acids.