Navigation Links
High-Level Expression of Peanut Allergens Affected by Rare Codon Usage

series (Novagen) that uses the T7 RNA polymerase responsive promoter to produce (His)10 -tagged fusion proteins. Two E. coli strains were transformed with this plasmid and selected on LB broth supplemented with ampicillin: BL21-Gold(DE3) cells and the recently generated BL21-CodonPlus (DE3)-RIL cells 6 carrying extra copies of the argU, ileY, and leuW tRNA genes. Recombinant Ara h 2 expression was induced by adding 1 mM IPTG when the cells reached an optical densitiy of 0.6 to 0.8 at 600 nm. Incubation was continued at three different temperatures (22C, 30C, 37C) for an additional 2, 4, and 20 hours.

The production of Ara h 2 was enhanced significantly with the new BL21-Codon Plus (DE3)-RIL cells (Figure 1C), compared to the pQE/M15 system (Figure 1A) and to the conventional BL21-Gold(DE3) cell expression system (Figure 1B). The highest amount of recombinant protein was achieved by selecting an overnight incubation at 22C after IPTG induction.

The final yield of soluble recombinant Ara h 2 was estimated as 5 mg/L of culture, which is 100 times greater when compared to the 50 g achieved by the pQE/M15 expression system.

Conclusions

It is possible to make large quantities of high-quality heterologous proteins that are otherwise difficult to express in conventional E.coli hosts. When low-expression yields or low-quality recombinant protein is due to rare codon usage, BL21-CodonPlus (DE3)-RIL cells are the hosts of choice. We used these special cells and greatly increased the expression yield of the major peanut allergen Ara h 2, which contains 12% of the rare AGG/AGA codons in its message.

Methods

A 10-ml culture of each of the expression experiments was induced by adding 1 mM IPTG, then incubation commenced at three different temperatures (22C, 30C,
'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence Humanized Renilla GFP Reporter
2. High-Level Protein Expression, One-Column Purification, and FLAG Epitope Tagging in E. coli
3. An Epitope Tagging Vector for Gene Expression in Mammalian Cells
4. Antibodies for Studying NMDA Receptor Protein Expression and Synapse-Specific Immunolabeling
5. Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
6. Innovative Tissue Array Technology for High-Throughput Screening of Gene Expression
7. Functional Cloning Using ViraPort Retroviral cDNA Expression Libraries
8. A New Lambda Vector for Mammalian Expression
9. Expression and Purification of Recombinant Proteins That Have Native Amino Acid Sequence
10. Codon Bias-Adjusted BL21 Derivatives for Protein Expression
11. Tools for Detecting MSH2 Expression in Chinese Hamster Ovary Cells
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: High Level Expression Peanut Allergens Affected Rare Codon Usage

(Date:9/30/2014)... Proove Biosciences , the commercial and research leader in ... test built upon research from their published P.A.I.N. Study. ... physicians with an objective way to understand theirs patient’s ... individuals. With this new test, physicians will be able ... that will affect their pain tolerance. , The P.A.I.N. ...
(Date:9/30/2014)... , September 30, 2014 ... browsing of chemistry in documents ... solutions and consulting services for life science research, ... and generate Markush structures from documents. ... intellectual property, however generating and understanding Markush structures ...
(Date:9/30/2014)... and ROCKVILLE, Md., Sept. 30, 2014 BioHealth ... on commercializing market-relevant biohealth innovations and increasing access to ... announced today that venture capitalist, Tania Fernandez , ... advisor. Dr. Fernandez will be a member of the ... will provide up to $50 million in seed and ...
(Date:9/30/2014)... September 30, 2014 The custom stainless ... Boston Product Show 2014. The Boston Area Chapter of ... 23rd annual, day-long event for Wed., Oct. 1 at ... to be the largest in ISPE Boston’s long history, ... of 3,000 attendees expected. , HOLLOWAY AMERICA President ...
Breaking Biology Technology:Proove Biosciences is Now Offering the Proove Pain Perception Genetic Test to Help Physicians Assess Patient Pain Tolerance Levels 2Proove Biosciences is Now Offering the Proove Pain Perception Genetic Test to Help Physicians Assess Patient Pain Tolerance Levels 3ChemAxon Introduces ChemCurator for Chemical Information Extraction and Markush Structure Creation 2Seasoned Life Sciences Venture Capitalist Tania Fernandez Joins BHI to Help Manage New BioHealth Gap Fund 2HOLLOWAY AMERICA to Exhibit at ISPE Boston Product Show 2014 2HOLLOWAY AMERICA to Exhibit at ISPE Boston Product Show 2014 3
... Contract Research Organization Exceeds Expectations, MEMPHIS, ... Contract Research,Organization exclusively focused on oncology, announced ... exceeded projections as the company continues,to experience ... to,ACORN,s ability to consistently exceed client expectations, ...
... Pharma, Inc., a,diversified specialty pharmaceutical company, announced ... issues in the treatment of moderate to,severe ... dry eye medication Lacrisert(R),(hydroxypropyl cellulose ophthalmic insert), ... helps to retain,moisture, stabilize the tear film, ...
... -- 3SBio Inc. (Nasdaq:,SSRX), a leading biotechnology company ... primarily in China,today announced that it has filed ... for approval of NuLeusin for the treatment of ... is expected to,be the only treatment of this ...
Cached Biology Technology:ACORN CRO Continues Growth in Strong Third Quarter 2Dry Eye Survey: Ophthalmologists Cite Treatment Limitations/Gaps in Treatment of Moderate to Severe Dry Eye 2Dry Eye Survey: Ophthalmologists Cite Treatment Limitations/Gaps in Treatment of Moderate to Severe Dry Eye 3Dry Eye Survey: Ophthalmologists Cite Treatment Limitations/Gaps in Treatment of Moderate to Severe Dry Eye 4Dry Eye Survey: Ophthalmologists Cite Treatment Limitations/Gaps in Treatment of Moderate to Severe Dry Eye 53SBio Inc. Files for SFDA Approval of NuLeusin 23SBio Inc. Files for SFDA Approval of NuLeusin 33SBio Inc. Files for SFDA Approval of NuLeusin 4
(Date:9/30/2014)... you want your doctor to know what goes wrong with ... good idea to know what "normal" actually is. That,s where ... of the FASEB Journal comes in. ... transcriptomea key set of molecules that can help scientists "see" ... time. What,s more, they found never-before-detected gene activity and that ...
(Date:9/30/2014)... in German . ... to risky situations such as exposure to predators. Researchers ... in a long-term study on different populations of great ... and ambient temperature. High metabolic rates and low temperatures ... scenarios birds were more likely to approach potential predators. ...
(Date:9/30/2014)... perilous threats in today,s ocean. From overfishing and ... coral ecosystems are disappearing at unprecedented rates around ... corals surrounding the island of Moorea in French ... environment: coral guard-crabs. New research from the National ... Seabird McKeon and the museum,s predoctoral fellow Jenna ...
Breaking Biology News(10 mins):Scientists identify which genes are active in muscles of men and women 2Risky metabolism 2Smithsonian scientists discover coral's best defender against an army of sea stars 2
... A cell,s genome maintains its integrity by organizing ... DNA called heterochromatin. In the comparatively simple organism fission ... plays an essential role in assembling heterochromatin, which keeps ... Central to this process is a large protein complex ...
... of vitamin C were associated with higher levels of high ... issues or death for heart failure patients, in research presented ... to those with high vitamin C intake from food, heart ... intake were 2.4 times more likely to have higher levels ...
... The hermaphroditic cleaner shrimp Lysmata amboinensis usually ... structure. New research published in BioMed Central,s open access journal ... any group larger than two, viciously attack and kill each ... amboinensis are protandric simultaneous hermaphrodites. This means that they ...
Cached Biology News:CSHL team solves a protein complex's molecular structure to explain its role in gene silencing 2Low vitamin C levels may raise heart failure patients' risk 2Violent passions -- jealous cleaner shrimp murder their rivals 2
All serum is collected from U.S.D.A. fryers and broilers. Pricing: $15/unit for 1 - 10 units; $12/unit for 11 - 50 units...
RAT ANTI MOUSE TCR V BETA 7:RPE Immunogen: T-cell lymphoma TK1...
ANTI S. ENT TOX BC2D...
... Anti-Major DNA-binding protein(DBP). Protein ... NP_001343 ELISA titre using ... Suggested conditions for Western Blot: ... buffer, and HRP conjugated anti-Rabbit IgG should ...
Biology Products: