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

s of the S. pombe host strain and reagents for protein induction and purification.

The pESP-1 Expression Vector

figure 1

Designed for expression in S. pombe, the pESP-1 vector (figure 1) is composed of the following parts: (1) a ColE1 origin of replication and an ampicillin resistance (Ampr) fragment, which allow vector replication and antibiotic selection in E. coli; (2) the ars1 fragment, which provides an origin of replication for the vector to replicate autonomously in S. pombe; (3) a LEU2-d gene from Saccharomyces cerevisiae, which serves as a selection marker for transforming the expression vector into S. pombe cells (strain SPQ01) and (4) the expression cassette, which contains the S. pombe nmt1 promoter, a translational start site, a GST protein-tag sequence, a multiple cloning site and the nmt1 transcription termination signal. Insertion of the gene of interest into the multiple cloning site results in an N-terminal fusion with the GST peptide, which facilitates one-step purification of the GST fusion protein.2

Figure 3

The nmt1 promoter of S. pombe is tightly repressed in the presence of thiamine (vitamin B1) in the growth medium and is highly activated upon its removal.3 When activated, the nmt1 promoter has been shown to be one of the strongest promoters in S. pombe.4 For this reason, the nmt1 promoter has been widely used to study gene function in S. pombe. Figure 3 shows Western blot analysis of the GST-calmodulin fusion protein in the pESP-1 vector that has been expressed in cells grown under
'"/>

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:7/10/2014)... July 10, 2014   Ceres, Inc . (Nasdaq: ... company, announced today financial results for the three months ... its business. Ceres reported that the company ... performance this growing season in Brazil, which concluded in ... company,s sorghum evaluation areas for part of the growing ...
(Date:7/10/2014)... June 17, 2014 Using microscopic polymer light ... gases, researchers at MIT,s Quantum Photonics Laboratory have ... in the parts-per-billion range. Optical sensors are ideal ... high signal-to-noise ratio, compact, lightweight nature, and immunity ... sensors had been developed before, the MIT team ...
(Date:7/10/2014)... researchers have observed the fractional quantum Hall effect in ... matter can be tuned by an electric field. ... when electrons confined to thin sheets are exposed to ... behavior where thousands of individual electrons behave as a ... effect is well established, many details of this collective ...
(Date:7/10/2014)... theatergoers have shelled out for crude 3-D glasses, ... viewing experience. These basic devices, used to trick ... may soon be rendered obsolete with the introduction ... University researchers. , TAU doctoral students Yuval Yifat, ... efficient holography based on nanoantennas that could be ...
Breaking Biology Technology:Ceres Announces Fiscal Third Quarter 2014 Financial Results 2Ceres Announces Fiscal Third Quarter 2014 Financial Results 3Ceres Announces Fiscal Third Quarter 2014 Financial Results 4Ceres Announces Fiscal Third Quarter 2014 Financial Results 5Ceres Announces Fiscal Third Quarter 2014 Financial Results 6Ceres Announces Fiscal Third Quarter 2014 Financial Results 7Ceres Announces Fiscal Third Quarter 2014 Financial Results 8Ceres Announces Fiscal Third Quarter 2014 Financial Results 9Swell new sensors 2Columbia researchers observe tunable quantum behavior in bilayer graphene 2Columbia researchers observe tunable quantum behavior in bilayer graphene 3Projecting a 3-dimensional future 2Projecting a 3-dimensional future 3
... ... People, INDIANAPOLIS, Oct. ... it has made the list of Top 20,biopharmaceutical employers, according to the ... in the biotechnology, biopharmaceutical, pharmaceutical and,related industries. This year Lilly ranks number ...
... 10 EntreMed, Inc.,(Nasdaq: ENMD ) a ... cancer and inflammatory diseases,announced today that Thomas H. ... Development, will present a Company,overview at the 16th ... 12-14, 2008 in London, United Kingdom. Mr. Bliss, ...
... dry adhesive got a new competitor this week with a ... by a team of researchers from four U.S. institutions. , ... lizards to scurry up walls and cling to ceilings by ... microscopic branched elastic hairs in their toes that take advantage ...
Cached Biology Technology:Lilly Recognized as One of the World's Top Biopharmaceutical Employers 2EntreMed to Present at BioPartnering Europe Conference 2EntreMed to Present at BioPartnering Europe Conference 3Dry adhesive based on carbon nanotubes gets stronger, with directional gripping ability 2Dry adhesive based on carbon nanotubes gets stronger, with directional gripping ability 3Dry adhesive based on carbon nanotubes gets stronger, with directional gripping ability 4
(Date:7/10/2014)... Sweden , July 7, 2014 ... its first major design win (DW). A Chinese Top 5 ... the planned start of mass production in October 2014.   ... a Chinese Top 5 smartphone OEM, which has a planned ... and a target sales volume for this phone of 3 ...
(Date:7/10/2014)... July 8, 2014 Fingerprint ... new design win (DW). An Asian OEM has selected FPC1021 ... production start in August 2014.   FPC is ... which has a planned date for start of mass production ... order of SEK 5M, for delivery in the first half ...
(Date:7/10/2014)... , July 9, 2014 The widespread ... in large part by meaningful use and related ... providers interact with laboratory information. Now, more than ... field of clinical informatics in order to deliver ... To address the educational needs of pathologists in ...
Breaking Biology News(10 mins):FPC Wins DW for 1021 Touch Sensor for Flagship Smartphone from Chinese Top 5 Smartphone OEM 2FPC Wins DW and Initial Ramp Order for FPC1021 Touch Sensor for Flagship Smartphone from Asian OEM 2New Clinical Informatics Curriculum Prepares Future Pathologists 2New Clinical Informatics Curriculum Prepares Future Pathologists 3New Clinical Informatics Curriculum Prepares Future Pathologists 4
... child-resistant packaging for most medications has contributed to the ... 500,000 calls are made to poison control centers each ... and the number of emergency department visits for unsupervised ... scheduled for publication in The Journal of Pediatrics ...
... into the evolution of microbial communities, an ideal opportunity ... become established and how ecosystems begin to form. Soils ... with time under the influence of multiple environmental factors ... advancements in the science and management of soils. ...
... Chicago,s Field Museum and international collaborators have described a new ... bizarre lower spine on Earth. The interlocking vertebrae of ... more robust relative to body mass, a condition not found ... enigma to evolutionary biologists, with no known adaptive significance. ...
Cached Biology News:Flow restrictors may reduce young children's accidental ingestion of liquid medications 2University of Alberta scientists get dirty at the Robson Glacier 2New species of Hero Shrew found in equatorial Africa 2
... Lipid adsorption and clarification reagent; ... the purification of antibodies, proteins, nucleic ... has negligible cross-reactivity, and improves downstream ... a solid-phase, non-ionic adsorbent supplied as ...
A simple, sensitive and reliable method for determining free sulfhydryl content in peptides, protein...
... Freedom EVO Clinical is Tecans open platform ... enhance quality, safety and performance in the ... is provided by flexible barcode reading and ... mention that the system complies with the ...
... Conjugate Stabilizer is an aqueous solution that ... non-toxic stabilizing chemicals in a MOPS buffer ... This product contains a combination of ... Proclin 300 (Rohm and Haas Company) as ...
Biology Products: