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

ides sufficient thiamine to repress the nmt1 promoter. Expression is induced by harvesting the cells and growing them in EMM broth without thiamine for approximately 18 hours.

Figure 2

The chicken calmodulin gene was inserted into the pESP-1 vector and expressed in S. pombe. Figure 2 shows the cell lysate samples derived from repressed and induced cells containing chicken calmodulin. When cells were grown under induced conditions, the crude lysate shows a dominant band at the size expected for a GST-calmodulin fusion protein (43 kDa) (figure2, lane 4). In the lysate of cells grown under repressed conditions, this band was absent. (Figure 2, lane 1) When the samples were subjected to one-step purification of the GST fusion protein by using GST affinity resin, the induced GST-calmodulin fusion protein was depleted in the glutathione resin flowthrough sample (Figure 2, lane 5) and was eluted to near homogeneity in the glutathione elution fraction (Figure 2, lane 6). No detectable GST fusion protein was observed when lysate derived from repressed cells was subjected to the same purification procedure (Figure 2, lane 3). Figure 3 shows Western blot analysis of the GST-calmodulin fusion protein in cell lysates derived from repressed and induced cells. The fusion protein was not detected in cells when the promoter was repressed but was abundant in cells when the promoter was induced. We have also observed that constitutive, high-level expression of GST fusion proteins is possible by inoculating the cells directly into the induction medium (EMM), which significantly simplifies the overall procedure. However, proteins
'"/>

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:9/17/2014)... , September 17, 2014 ... of action from a truly novel class ... MGB Biopharma, a biopharmaceutical company developing ... today that it has secured £4.0m ($6.4m) ... antibacterial, MGB-BP-3, against a range of Gram-positive ...
(Date:9/16/2014)... Sept. 16, 2014 This report analyzes the worldwide ... Segments: Software, Hardware, and Biocontent. The report provides separate comprehensive ... Japan , Europe , ... and Rest of World. Annual estimates and forecasts are provided ... analysis is provided for these markets. Market data and analytics ...
(Date:9/16/2014)... University scientists who created a deicing film for radar ... a transparent coating for glass. , The new ... could keep glass surfaces from windshields to skyscrapers free ... radio frequencies (RF). , The technology was introduced ... Applied Materials and Interfaces . , The material ...
(Date:9/16/2014)...  The Council for Entrepreneurial Development (CED), the ... country, today released a mid-year update to its ... It showed that in the first half of ... in the technology, life science, advanced manufacturing and ... variety of sources. Significant investment in technology companies and ...
Breaking Biology Technology:MGB Biopharma Secures £4.0m ($6.4m) to Advance its Novel Lead Antibacterial, MGB-BP-3, into Clinical Development 2MGB Biopharma Secures £4.0m ($6.4m) to Advance its Novel Lead Antibacterial, MGB-BP-3, into Clinical Development 3MGB Biopharma Secures £4.0m ($6.4m) to Advance its Novel Lead Antibacterial, MGB-BP-3, into Clinical Development 4MGB Biopharma Secures £4.0m ($6.4m) to Advance its Novel Lead Antibacterial, MGB-BP-3, into Clinical Development 5MGB Biopharma Secures £4.0m ($6.4m) to Advance its Novel Lead Antibacterial, MGB-BP-3, into Clinical Development 6MGB Biopharma Secures £4.0m ($6.4m) to Advance its Novel Lead Antibacterial, MGB-BP-3, into Clinical Development 7Global Bioinformatics Industry 2Global Bioinformatics Industry 3Global Bioinformatics Industry 4Global Bioinformatics Industry 5Global Bioinformatics Industry 6Global Bioinformatics Industry 7Global Bioinformatics Industry 8Global Bioinformatics Industry 9Global Bioinformatics Industry 10Global Bioinformatics Industry 11Global Bioinformatics Industry 12Global Bioinformatics Industry 13Global Bioinformatics Industry 14Global Bioinformatics Industry 15Global Bioinformatics Industry 16Global Bioinformatics Industry 17Global Bioinformatics Industry 18Global Bioinformatics Industry 19Global Bioinformatics Industry 20Global Bioinformatics Industry 21Global Bioinformatics Industry 22Global Bioinformatics Industry 23Nanoribbon film keeps glass ice-free 2CED's Mid-Year Update to its Innovators Report Shows NC Startups Attracting Investment From Across the Globe with Several High-Profile Investment Deals, IPOs and Acquisitions Signaling Strength of Entrepreneurial Sector 2CED's Mid-Year Update to its Innovators Report Shows NC Startups Attracting Investment From Across the Globe with Several High-Profile Investment Deals, IPOs and Acquisitions Signaling Strength of Entrepreneurial Sector 3CED's Mid-Year Update to its Innovators Report Shows NC Startups Attracting Investment From Across the Globe with Several High-Profile Investment Deals, IPOs and Acquisitions Signaling Strength of Entrepreneurial Sector 4
... Mylan Inc. (NYSE: MYL ),today announced that Mylan ... and Drug Administration (FDA) for its,Abbreviated New Drug Application ... and 200 mg., Lamotrigine Tablets are the generic ... approximately $1.97 billion,for the 12 months ending Sept. 30, ...
... Calif., Dec. 12 LUMEDX Corporation, the ... and imaging,solutions, introduces CardioManager Performance Management System(TM),cardiology ... operational,financial -- into accessible, actionable information. CardioManager,s,performance ... a,high-level view of data and the ability ...
... the FDA for the Fourth Genmab Antibody ... Collaboration with Roche., COPENHAGEN, December 12 Genmab ... application (IND) for a,Genmab antibody developed under the company,s ... Roche. Genentech and Roche are collaborating on,development of the ...
Cached Biology Technology:New Cardiology Software Helps Heart Centers Manage for Optimal Clinical Performance 2Fourth Genmab Antibody Developed Under Roche Collaboration to Enter Clinic 2Fourth Genmab Antibody Developed Under Roche Collaboration to Enter Clinic 3
(Date:9/16/2014)... have discovered the "molecular brakes" that time the generation ... mice. These "hair cells" translate sound waves into electrical ... interpreted as sounds. If the arrangement of the cells ... of the research will be published in The ... proteins Hey1 and Hey2 act as brakes to prevent ...
(Date:9/16/2014)... The meteorite impact that spelled doom for the dinosaurs ... flowering plants to a much greater extent than their ... researchers. The results are published in the journal ... a treasure trove of thousands of fossilized leaves of ... team was able to reconstruct the ecology of a ...
(Date:9/16/2014)... Hesperides vessel returned to Spain culminating the around the ... clear picture of how the global ocean works and ... pollutants from the atmosphere is not limited to coastal ... of the planet, and it is already affecting the ... week in CSIC Residence for Researchers in Barcelona, in ...
Breaking Biology News(10 mins):Hey1 and Hey2 ensure inner ear 'hair cells' are made at the right time, in the right place 2Meteorite that doomed dinosaurs remade forests 2Meteorite that doomed dinosaurs remade forests 3Meteorite that doomed dinosaurs remade forests 4The Malaspina Expedition confirms that pollution reaches even the most remote areas of the ocean 2The Malaspina Expedition confirms that pollution reaches even the most remote areas of the ocean 3
... seafood, the options are many and so are some of the ... healthy? Is it safe? Is it endangered? While there are many ... an iPhone app a group of researchers have found a ... then it is likely to be healthy to eat too," said ...
... are more common than ever before. According to the ... Americans suffers from an allergy from milder forms ... allergies which can lead to anaphylactic shock. ... an allergic reaction, the treatment is too limited, says ...
... Boulder, Colo., USA Two Geology studies focus on ... view of alpine denudation and the other studying and ... include fossil microatolls and sea level; the potential rupture ... Northwest and British Columbia; paleoclimate; and the relationship between ...
Cached Biology News:Study finds healthy seafood comes from sustainable fish 2What sets allergies in motion? 2What sets allergies in motion? 3Debris flows, landslides, fossil microatolls, paleo-seasonality, and carbonate ore deposits 2Debris flows, landslides, fossil microatolls, paleo-seasonality, and carbonate ore deposits 3Debris flows, landslides, fossil microatolls, paleo-seasonality, and carbonate ore deposits 4Debris flows, landslides, fossil microatolls, paleo-seasonality, and carbonate ore deposits 5Debris flows, landslides, fossil microatolls, paleo-seasonality, and carbonate ore deposits 6Debris flows, landslides, fossil microatolls, paleo-seasonality, and carbonate ore deposits 7Debris flows, landslides, fossil microatolls, paleo-seasonality, and carbonate ore deposits 8