Navigation Links
Expression and Purification of Recombinant Proteins That Have Native Amino,,,Acid Sequence

he pCAL-n-EK vector, a new addition to the Affinity system, contains the CBP coding sequence followed by the EK cleavage site. This configuration allows efficient cleavage of all sequences at the N terminus of the polypeptide of interest and produces proteins with native amino acid sequence.3 Protein coding sequences are cloned into the pCAL-n-EK vector by LIC, a high-efficiency cloning method that does not require ligation or restriction enzyme digestion. LIC creates a seamless junction4,5,6 between the EK cleavage site and the protein coding sequence. The protein sequence is efficiently cloned into the pCAL-n-EK vector, and protein expression is induced by the addition of isopropyl-thio-D-galactoside (IPTG). The fusion protein is purified in one step using the CBP affinity tag. If desired, the affinity tag can be removed by proteolytic treatment with Stratagene's recombinant Enterokinase, and the recombinant protein of interest can be recovered free of CBP-containing digestion products and EK.

Design of the pCAL-n-EK Vector

figure 1

The pCAL-n-EK vector (figure 1) is the fourth in a series of CBP affinity-tag vectors released by Stratagene.1 It is derived from the pET-11 vector series and contains the lacIq gene for expression of the Lac repressor protein and the hybrid T7 promoter for controlled expression of the inserted protein coding sequence.7,8 The vector is transformed into specialized E. coli strains, such as Epicurian Coli BL21(DE3), which contains an expression cassette for T7 RNA polymerase that is induced in the presence of IPTG, allowing tight control and high-level, induced expression of the inserted protein coding sequence. The promoter is followed by the 26-amino-acid
'"/>

Source:


Page: All 1 2 3 4 5 6 7 8

Related biology technology :

1. An Epitope Tagging Vector for Gene Expression in Mammalian Cells
2. Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
3. High-Level Expression of Peanut Allergens Affected by Rare Codon Usage
4. Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
5. Innovative Tissue Array Technology for High-Throughput Screening of Gene Expression
6. New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence Humanized Renilla GFP Reporter
7. Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
8. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
9. A New Lambda Vector for Mammalian Expression
10. Codon Bias-Adjusted BL21 Derivatives for Protein Expression
11. Tools for Detecting MSH2 Expression in Chinese Hamster Ovary Cells
Post Your Comments:
(Date:7/7/2015)... Summary GlobalData,s ... report is an essential source of information and ... company data, deal analysis, corporate strategy, and market ... the current and future growth drivers of the ... factors shaping and driving the gene therapy business, ...
(Date:7/7/2015)... ... 2015 , ... bioLytical Laboratories Inc. announced today that its ... presentation at the 8th Annual International AIDS Society (IAS) Conference on HIV Pathogenesis, ... from 1:00 pm to 2:00pm Pacific Standard Time at the poster discussion on ...
(Date:7/7/2015)... ... ... uBiome, the leading microbial genomics company, is offering talented researchers with big ... laboratory analysis. The goal is to fast-track a study with the greatest potential to ... dashboard, enabling them to view and compare the results of their study, as well ...
(Date:7/7/2015)... ... 07, 2015 , ... The Pistoia Alliance, a global, not-for-profit ... science R&D, has attracted a number of new members from across the industry. ... joining as a Core member, and DNAnexus and Tessella as Participating members. Genexyx ...
Breaking Biology Technology:PharmaSphere: Emerging Biotechnologies - Gene Therapy Market Analysis 2bioLytical to Present Findings of Early HIV Detection 2bioLytical to Present Findings of Early HIV Detection 3uBiome to Award Grant of up to $100k for Groundbreaking Microbiome Research 2uBiome to Award Grant of up to $100k for Groundbreaking Microbiome Research 3Pistoia Alliance Continues to Attract New Members 2
... Cord-blood-derived Stem Cell Treatments -- SHENZHEN, China, Sept. 2 /PRNewswire-Asia/ -- ... ... ... var first_result; // Results are keyed by longUrl, so we need to grab the first one. for ...
... soon be used to convert metallic wastes into high-value ... the September issue of Microbiology . Researchers ... Birmingham have discovered the mechanisms that allow the common ... precious metal palladium from industrial waste sources. Palladium ...
... National Institute of Standards and Technology (NIST) have demonstrated ... data for a wide variety of fields dealing with ... as a few nanoliters. Currently a table-top prototype, the ... biotechnologists studying minute quantities of complex materials that must ...
Cached Biology Technology:New Research Demonstrates Safety of Cord-blood-derived Stem Cell Treatments 2New Research Demonstrates Safety of Cord-blood-derived Stem Cell Treatments 3New Research Demonstrates Safety of Cord-blood-derived Stem Cell Treatments 4New Research Demonstrates Safety of Cord-blood-derived Stem Cell Treatments 5New Research Demonstrates Safety of Cord-blood-derived Stem Cell Treatments 6New Research Demonstrates Safety of Cord-blood-derived Stem Cell Treatments 7New Research Demonstrates Safety of Cord-blood-derived Stem Cell Treatments 8Metal-mining bacteria are green chemists 2Micro rheometer is latest Lab On a Chip device 2
(Date:6/18/2015)... OXFORD, Conn. , June 18, 2015 NXT-ID, ... biometric authentication company focused on the growing mobile commerce market ... of a featured segment on "Money on the Mark", scheduled ... York City June 20 th . ... interview will be from 7:00 to 8:00pm EST. NXT-ID, Inc.,s ...
(Date:6/17/2015)... Germany , June 17, 2015 ... today launched new Investigator ® STR assay kits for ... the United States . The new genetic fingerprint kits ... markers (short tandem repeats or STRs) for DNA matching. They ... quality of DNA in each sample, a novel QIAGEN technology ...
(Date:6/17/2015)... June 17, 2015  Crossmatch™, a leading provider ... its U.are.U ® 4500 fingerprint readers have ... Eateries, Inc., a San Jose, CA ... increase security, improve accountability and reduce fraud at ... non-repudiable identity confirmation for employee sign in and ...
Breaking Biology News(10 mins):NXT-ID, Inc.'s CEO Gino Pereira Talks About the Wocket Smart Wallet on WABC Radio, New York City, June 20th 2NXT-ID, Inc.'s CEO Gino Pereira Talks About the Wocket Smart Wallet on WABC Radio, New York City, June 20th 3QIAGEN Launches New Genetic Fingerprinting Kits for U.S. Forensic Labs 2QIAGEN Launches New Genetic Fingerprinting Kits for U.S. Forensic Labs 3QIAGEN Launches New Genetic Fingerprinting Kits for U.S. Forensic Labs 4QIAGEN Launches New Genetic Fingerprinting Kits for U.S. Forensic Labs 5Togo's Eateries Improve Security and Accountability with Crossmatch Fingerprint Readers 2Togo's Eateries Improve Security and Accountability with Crossmatch Fingerprint Readers 3
... surrounds the DNA of our cells could help develop treatments ... have discovered that the proteins within this coating known ... different organs of the body. This variation ... proteins in the protective seal to cause illness in some ...
... Daejeon, Republic of Korea, February 26, 2013The Korea Advanced ... a global energy and petrochemicals enterprise, signed on February ... Agreement) on joint collaborations in research and development of ... Agreement was subsequently concluded upon the signing of the ...
... is faced with some of the world,s worst pollution. Until ... a major contributor to that pollution human-caused nitrogen emissions ... biology professor and Stanford Woods Institute senior fellow Peter Vitousek ... deposited on land and water in China by way of ...
Cached Biology News:Cell discovery could hold key to causes of inherited diseases 2Launched the Saudi Aramco-KAIST CO2 Management Center in Korea 2Stanford scientists help shed light on key component of China's pollution problem 2
... a magnetic bead-based PCR purification ... transfer, centrifugation or filtration steps. ... large PCR extension products in ... AMPure can easily be performed ...
... is designed for the fast and simple ... at any position (for the generation of ... is designed to allow neomycin / kanamycin ... combines a prokaryotic promoter (gb2) for expression ...
Includes a choice of any 3 EasySep mouse selection kits and EasySep magnet...
... trypsinizing flasks are used for ... cell suspensions by digestion of ... accepts cotton plugs. Four extra-deep ... Height of Baffles mm ...
Biology Products: