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:8/22/2014)... Greenville, SC (PRWEB) August 22, 2014 ... that indisputable acknowledgement is a national disgrace. Furthermore, ... part in establishing the current, deplorable state of ... outlined "5 great mistakes," to merely serve as ... convey the extent of colossal failure(s) that have ...
(Date:8/21/2014)... (PRWEB) August 21, 2014 ... Fertilizer Industry” is a professional and in-depth ... The report provides basic Seaweed Fertilizer information, ... industry chain structure as well as industry ... analysis, including domestic market as well as ...
(Date:8/21/2014)... BURLINGTON, Mass. , Aug. 21, 2014 /PRNewswire/ ... in Brazil and ... restrictions. The uptake of several newly approved agents, ... for this indication and the launch of emerging ... asthma markets. In particular, the emerging IL-5 inhibitors ...
(Date:8/21/2014)... Chester, NJ (PRWEB) August 21, 2014 ... of personal selling optimization technology and services for ... publication of “Grading Pharma’s Use of New Commercial ... of Measurement & Analytics. , The article examines ... marketing yield that are being tested in the ...
Breaking Biology Technology:5 Great Mistakes in Healthcare and How to Fix Them by Dr. Ira Williams 2Internationals Seaweed Fertilizer Industry 2014 Development Trend Analysis in New Research Study at DeepResearchReports.com 2Internationals Seaweed Fertilizer Industry 2014 Development Trend Analysis in New Research Study at DeepResearchReports.com 3Asthma Agents Currently Face a Suboptimal Coverage Environment in Brazil and Mexico, and as the Treatment Armamentarium Expands, Competitiveness Will Further Intensify 2Asthma Agents Currently Face a Suboptimal Coverage Environment in Brazil and Mexico, and as the Treatment Armamentarium Expands, Competitiveness Will Further Intensify 3Pursuit's Peter Robinson Publishes "Grading Pharma's Use of New Commercial Sales Models" 2
... -- Bruker AXS Inc. on Thursday announced the promotions ... to executive vice presidents, responsible for leading all aspects ... the Madison headquarters of Bruker AXS, now leads the ... as chief technology officer heads global R&D. , ,Burgaezy, ...
... describes today's go-it-alone economic climate than the word "entrepreneur." ... appealing to those who would strike out on their ... large and small along with colleges and universities and ... to help them in their quests to be successful ...
... CHICAGO -Last week, we talked about the size and ... key market segments. This week's column will focus on the ... , ,Before we comment on some trends from this ... absent here is the major part of the diagnostics business ...
Cached Biology Technology:Beware -- Use of the word 'entrepreneur' can put you in a legal mess 2Beware -- Use of the word 'entrepreneur' can put you in a legal mess 3Beware -- Use of the word 'entrepreneur' can put you in a legal mess 4Beware -- Use of the word 'entrepreneur' can put you in a legal mess 5Midwest plays key role in developing new medical device companies 2Midwest plays key role in developing new medical device companies 3
(Date:8/21/2014)... is a key region in the global oceanic circulation. "On ... warm Atlantic water flows to the north into the Arctic ... and sea ice push their way out of the Arctic ... water cools here on its way to the north and ... in this manner drives the global band of oceanic currents ...
(Date:8/21/2014)... La. A team of researchers at ... method for using affordable, consumer-grade 3D printers ... that can contain antibacterial and chemotherapeutic compounds ... comprised of doctoral students and research faculty ... engineering programs collaborated to create filament extruders ...
(Date:8/20/2014)... In a study of 158 pregnant teenagers in ... craving and intentional consumption of ice, cornstarch, vacuum dust, ... a new Cornell study. , Moreover, such teens had ... did not eat nonfood substances. , Pregnant teens, regardless ... which can lead to iron deficiency and anemia. Low ...
Breaking Biology News(10 mins):Arctic sea ice influenced force of the Gulf Stream 2Arctic sea ice influenced force of the Gulf Stream 3Louisiana Tech University researchers use 3D printers to create custom medical implants 2Louisiana Tech University researchers use 3D printers to create custom medical implants 3Pica in pregnant teens linked to low iron 2
... explain the complexities of the origins of life. Their work, ... Interface , offers new insights into how RNA signaling likely ... that today,s genetic code probably resulted from a combination of ... student at NYU School of Medicine and the paper,s lead ...
... When a woman becomes pregnant, she knows it is important ... of herself and the health of her baby. New clinical ... Eurpean Federation of Periodontology (EFP) urge pregnant women to maintain ... periodontal disease may be at risk of adverse pregnancy outcomes, ...
... role of microbiology in agriculture combined with new technologies ... population increases according to a new report from the ... are essential partners in all aspects of plant physiology, ... solely on the plant," says Ian Sanders of University ...
Cached Biology News:Researchers develop model of 'near-optimal' genetic code 2Expectant mothers' periodontal health vital to health of her baby 2Report proposes microbiology's grand challenge to help feed the world 2
sterile filtered, off the clot, all male donors, Total Protein: 4.5 to 8.5 g/dl...
Recombinant Mouse MMP-9, CF...
Human/Mouse/Rat JNK Pan Specific Affinity Purified PAb...
Blue Standard is 3-cyano-7-hydroxycoumarin....
Biology Products: