Navigation Links
Yeast Protein Production System Features High Yields and One-Step,,,Purification

repressed and induced conditions. The fusion protein cannot be detected in cells when the promoter is repressed and is abundant in cells when the promoter is induced. In addition to inducing expression of the gene product of interest, constitutive high-level expression is possible by growing the clones in media without thiamine.

The pESP-1 vector also contains DNA sequences coding for amino acid residues that are specifically recognized and cleaved by enterokinase and thrombin. These sites are located between the GST tag and the multiple cloning site (figure 1). Insertion into the multiple cloning site results in fusion proteins that can be detached from the GST tag with either protease. Cleavage by thrombin results in a fusion protein having the FLAG peptide (DYKDDDDK)5 at its N terminus; antibodies against the FLAG peptide are available from Stratagene.

Establishing the Expression Strain

The gene of interest is inserted into the multiple cloning site of the pESP-1 vector and is fused in frame to the upstream GST gene. Successful cloning can be confirmed by sequence analysis. To establish the expression strain, the verified clone is then transformed into the competent cells of S. pombe host strain SP-Q01 ((leu1-32h ) that are included in the kit. These competent cells provide optimal transformation conditions. Transformants can be identified by their ability to grow on minimal medium (EMM) agar plates supplemented with 25 M thiamine. The expression strain can be stored at 70C and used for multiple experiments.

Induction and Purification of GST-Tagged Fusion Proteins

To induce expression of the protein of interest, the S. pombe expression strain is first grown to mid-logarithmic phase in the vegetative growth medium (YES medium), which prov
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. Epitope-Tagging Vectors for Functional Analysis in Yeast
2. New Yeast Cloning System for Producing Proteins with Native Amino Acid Sequences
3. Electroporation of Yeast Artificial Chromosomes
4. New Automated Yeast Cell Counter Ends Tedium & Errors of Manual Counting
5. High Yield Protein Production from Pichia pastoris Yeast: A Protocol for Benchtop Fermentation
6. A New C-Terminal GST Vector for Protein Production in S. pombe
7. New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
8. Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
9. Simple, Sensitive, and Rapid Detection of FLAG -Tagged Fusion Proteins
10. Optimized Imaging of Protein Gels Stained with Coomassie Brilliant Blue Dye
11. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
Post Your Comments:
(Date:7/22/2014)... July 22, 2014 Automated cell culture ... maintenance of cell culture. They allow production of optimum ... of researchers in drug discovery and industries and biopharmaceutical ... mAb production, iPSC research, cryobanking, cell-based assays, and cellular ... risk. , Analysts forecast the Global Cell Culture ...
(Date:7/22/2014)... 22, 2014 Two new members were ... Directors at its annual Shareholders Meeting on June 13: ... of The PFM Group, and Stephen T. Zarrilli, President ... announced the election of Michael A. DiPiano, Managing General ... and Richard P. Jaffe, Esq., of Duane Morris LLP, ...
(Date:7/22/2014)... CAMBRIDGE, Massachusetts og SHENZHEN ... BGI Tech Solutions Co., LTD, (BGI Tech), et ... globale lancering af en ny human whole exome-sekvenseringstjeneste ... Complete Genomics, højt anset i branchen for ... og højfølsom registrering af alle typer varianter, blev ...
(Date:7/22/2014)... SHENZHEN, China , July 22, 2014 /PRNewswire/ ... a subsidiary of BGI, the world,s largest genomics ... new human whole exome sequencing service based on ... includes in-depth bioinformatics analysis and SNP validation, and ... 2014. Complete Genomics, highly regarded ...
Breaking Biology Technology:Global Cell Culture Market to Grow at 10.27% CAGR by 2018 Forecasts a New Research Report ReportsnReports.com 2Global Cell Culture Market to Grow at 10.27% CAGR by 2018 Forecasts a New Research Report ReportsnReports.com 3Global Cell Culture Market to Grow at 10.27% CAGR by 2018 Forecasts a New Research Report ReportsnReports.com 4Two New Members Elected to University City Science Center Board of Directors 2Two New Members Elected to University City Science Center Board of Directors 3BGI Tech lancerer Human Whole Exome-sekvenseringstjeneste på Complete Genomics avancerede platform 2BGI Tech Launches Human Whole Exome Sequencing Service on Complete Genomics' Advanced Platform 2BGI Tech Launches Human Whole Exome Sequencing Service on Complete Genomics' Advanced Platform 3
... receive funding for technology development projects from the ... Grant program. , ,The TAG program provides grants ... companies to offset the cost of professional services. ... Automated Notification Systems, Inc. , Eau Claire; Micrablate, ...
... the balkanization of information technology in healthcare will take ... Digital Healthcare Conference 2006 . And vendors aren't necessarily ... customers. , ,Providers are facing that common IT problem: ... of vendors and systems with challenges agreeing to standards, ...
... a gathering of health professionals attending the eHealth Summit ... first state to have a fully integrated electronic medical ... barriers. , ,Speaking before a group of healthcare executives, ... state's eHealth Initiative, Doyle said those barriers include organizational ...
Cached Biology Technology:Being proactive with vendors helps healthcare improve IT 2Doyle asks health professionals to overcome barriers 2Doyle asks health professionals to overcome barriers 3
(Date:7/22/2014)... cells offer much promise for treating damaged organs and ... is poor, limiting their effectiveness. New methods are being ... their therapeutic function after transplantation, as described in a ... peer-reviewed journal from Mary Ann Liebert, Inc., publishers. The ... Access website . , In the ...
(Date:7/22/2014)... July 22, 2014 BGI Tech Solutions Co., ... world,s largest genomics organization, announced today the global ... service based on Complete Genomics, industry-leading platform. This ... SNP validation, and is available at reduced prices ... highly regarded in the industry for providing proprietary ...
(Date:7/22/2014)... emerging as a world-famous vacation spot with natural tourism resources, ... island. The Korea Institute of Geoscience and Mineral Resources (KIGAM) ... recent volcanic eruption was evident 5,000 years ago. That is ... lava spewed out of a volcano 5,000 years ago in ... one the whole peninsula. , The research team led by ...
Breaking Biology News(10 mins):BGI Tech launches human whole exome sequencing service on complete genomics' advanced platform 2Jeju Island is a live volcano 2
... some amphibians possess a unique sensory capability in the ... "touch" objects in their surroundings without direct physical contact ... Hermmen and his team in the physics department of ... for this sensory system. What they discover might one ...
... A method to detect contaminants in municipal water supplies ... Laboratory researchers whose findings are published on line in ... demonstrates that the technology that uses algae as sentinels ... Miguel Rodriguez Jr. and Elias Greenbaum of the Department ...
... to the "hardwired for news" theory, people devote more ... test the theory, University of Missouri researchers examined the ... researchers found that news about local health threats increased ... distant, or non-local, health threats. "Although journalists ...
Cached Biology News:Mathematical keys to a sixth sense -- the lateral-line system 2Mathematical keys to a sixth sense -- the lateral-line system 3Mathematical keys to a sixth sense -- the lateral-line system 4ORNL scientists hone technique to safeguard water supplies 2Online news garners more attention from readers if it's negative and localized, MU study finds 2
MOUSE ANTI HUMAN MBP (67-74) Immunogen: Human myelin basic protein...
... is designed to purify double-stranded DNA (dsDNA) generated ... PCR. The process is simple and fast. DNA ... can be efficiently purified and concentrated. The DNAclear ... bound to the Microfilter Cartridge in the presence ...
Normal donkey serum collected from healthy normal donkeys....
Whole blood is collected from canines (any breed, either sex), refrigerated within 6 hours, processed within 24 hours and frozen at -20C....
Biology Products: