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

potentially toxic to the host cannot be overexpressed under constitutive expression conditions. Using the S. pombe expression system, we have purified 10 different proteins, including the rat c-Jun N-terminal kinase (JNK) and the human MEK1 gene products (figure 4). Yield for these proteins has varied from 1.0 mg per liter to 12.5 mg per liter. Some of these proteins, especially MEK1, had been difficult to produce in significant quantities in E. coli.

figure 4

Removing the GST Tag by Proteolytic Cleavage with Enterokinase or Thrombin

In many cases, GST tags do not interfere with the function of the proteins. Our functional assays indicated that the biological activities of the GST-calmodulin fusion protein and the GST-MEK1 fusion protein expressed in S. pombe were retained (data not shown). However, if desired, the GST tag can be removed by treating the purified fusion protein with either enterokinase or thrombin to cleave between the GST tag and the protein of interest (figure 1). Figure 5 shows an example of protease cleavage of a GST-MEK1 fusion protein. The GST tag was efficiently detached from the MEK1 moiety by digesting the purified GST-MEK1 fusion protein with enterokinase. The enterokinase site is often preferred for cleavage since the resulting product has fewer additional amino acids remaining at the N terminus. Alternatively, if the recombinant protein is cleaved with thrombin, the resulting polypeptide will contain the FLAG peptide at its N terminus, which could facilitate further analysis of the protein.


'"/>

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:10/25/2014)... (PRWEB) October 24, 2014 The ... the nation’s leading nonprofit authority on the use ... information exchange, announces its appointment of four new ... The workgroup leaders include:, ,     Peter ...     Erik Pupo, Specialist Leader, Deloitte Consulting LLP ...
(Date:10/25/2014)... (PRWEB) October 24, 2014 SonaCare ... focused ultrasound (HIFU) technology, announced that its Board ... as Chief Executive Officer, President and member of ... Dr. Carol will take-on expanded responsibilities within the ... as well as overseeing commercialization, manufacturing and finance. ...
(Date:10/25/2014)... , October 24, 2014 ... clinical-stage pharmaceutical company focused on the development of oral ... Nadav Kidron will present at FireRock Capital,s ... Capital,s Annual Micro-Cap Conference ,Date: Tuesday, October 28, 2014 ... New York City , ...
(Date:10/25/2014)... -- Investor-Edge has initiated coverage on the ... Vertex Pharmaceuticals Inc. (NASDAQ: VRTX ), Alexion ... Inc. (NASDAQ: PPHM ), and StemCells Inc. ... be accessed at: http://investor-edge.com/register . The ... a positive note as the Dow Jones Industrial Average ...
Breaking Biology Technology:WEDI Appoints Four New Security and Privacy Workgroup Co-chairs 2SonaCare Medical Board of Directors Appoints Dr. Mark Carol as CEO and President 2SonaCare Medical Board of Directors Appoints Dr. Mark Carol as CEO and President 3Oramed to Present at the FireRock Annual Micro-Cap Conference 2Oramed to Present at the FireRock Annual Micro-Cap Conference 3Biotech Stocks Technical Data - CytRx, Vertex Pharma, Alexion Pharma, Peregrine Pharma, and StemCells 2Biotech Stocks Technical Data - CytRx, Vertex Pharma, Alexion Pharma, Peregrine Pharma, and StemCells 3Biotech Stocks Technical Data - CytRx, Vertex Pharma, Alexion Pharma, Peregrine Pharma, and StemCells 4Biotech Stocks Technical Data - CytRx, Vertex Pharma, Alexion Pharma, Peregrine Pharma, and StemCells 5
... for Combining Efficiency and Renewables into an Energy ... recognized as an important component to America' energy ... discussion about including Energy Efficiency into what many ... Standards (RPS) . The whole idea being that ...
... Merge eFilm announced Wednesday that its net income ... same quarter last year. Though its net sales, at ... bottom line suffered because it paid more than twice ... quarter ending June 30 were $1,497,000, up from $1,400,000 ...
... partnership to provide travel solution , ,ROCHESTER, ... Concur Technologies, Inc. announced Monday the formation ... that will deliver benchmarking information to travel managers ... Runzheimer, an international consulting firm specializing in travel ...
Cached Biology Technology:Energy efficiency is no longer the Rodney Dangerfield of energy 2Energy efficiency is no longer the Rodney Dangerfield of energy 3Runzheimer and Concur develop travel management service 2
(Date:10/15/2014)... from strains of influenza virus increases with the latest ... complacent that the most substantial threats have been identified, ... arise when a new virus strain – against which ... in the human population. There have been five such ... which – the 1918 Spanish Flu – cost 50 ...
(Date:10/15/2014)... NXT-ID, Inc. (NASDAQ: NXTD and NXTDW) ... growing mobile commerce market releases photos and video of the recent ... October 13 th . Gino Pereira , ... angel investor Mr. Chad A. Verdi rang the bell.  ... his investors and employees "for their work and dedication in bringing ...
(Date:10/14/2014)... of years since T. rex took its ... scientists is breathing life back into dinosaurs using high-powered ... research has important implications for how dinosaurs used their ... sense of smell and cool their brains. , ... student Jason Bourke, lead author of the new study ...
Breaking Biology News(10 mins):Scientists 'must not become complacent' when assessing pandemic threat from flu viruses 2Photo Release: NXT-ID Inc. Rings Opening Bell of the NASDAQ Stock Market October 13th 2Photo Release: NXT-ID Inc. Rings Opening Bell of the NASDAQ Stock Market October 13th 3Dinosaur breathing study shows that noses enhanced smelling and cooled brain 2Dinosaur breathing study shows that noses enhanced smelling and cooled brain 3
... -- Scientists from The Forsyth Institute and Kings College, ... community, the Human Oral Microbiome Database, which provides descriptions ... commonly found in peoples mouths. Most bacteria living ... maintaining the health of teeth and gums, as well ...
... a biosensing nanodevice developed by Arizona State University ... airport security checkpoints and revolutionize health screenings for ... aureus (MRSA). Even more incredible than the ... the worlds tiniest rotary motor: a biological engine ...
... ... covert data -, LAKE WORTH, Fla., March 24 ... produce the,world,s first self-authenticating passport. It was introduced in the new,Belgian passport ... produces the Belgian passport,has added an optical decoder page in the passport ...
Cached Biology News:Scientists launch human oral microbiome database 2Biosensing nanodevice to revolutionize health screenings 2Biosensing nanodevice to revolutionize health screenings 3FCO and GSSC Team to Create World's First Self-Authenticating Passport 2
... work in collaboration with Pall Life Sciences ... in a variety of membrane chemistries. The ... on the Caliper TPWII, MultiDose, Prelude, and ... our Automation Certified Syringe Filters are available ...
... The Mini-PROTEAN 3 Dodeca cell with multi-casting ... of up to 12 mini format handcast (8.3 ... cm) gels. The cell requires 3.4-4.4 L of ... cooling coil that attaches to an external refrigerated ...
...
... has been developed to significantly increase the ... Stability is increased at room temperature ... This product is subjected to 0.2 ... azide. This formulation insures consistently high ...
Biology Products: