Navigation Links
Generate Adenovirus Vectors in E. coli by Homologous Recombination,,,with the AdEasy Adenoviral Vector System

combination in E. Coli BJ5183

The AdEasy system is unique to other adenoviral vector systems because homologous recombination between shuttle vector and adenovirus genome is performed in E. coli. The E. coli strain BJ5183 was selected for its recombination capabilities and higher transformation efficiency. To validate production of recombinant adenoviral plasmids in BJ5183, the shuttle vectors described in the previous section were linearized with Pme I, dephosphorylated, and gel purified. Linear shuttle vectors were then electrotransformed into BJ5183 cells, both alone to serve as controls, and cotransformed with the pAdEasy-1 vector to generate recombinants. Miniprep DNA was prepared from picked kanamycin-resistant colonies and analyzed by digestion with Pac I. Positive recombinants were identified by the presence of a large fragment of approximately 30 kb and a smaller fragment of either 3.0 kb or 4.5 kb (depending on whether recombination occurred at the left arm or the origin of replication). Representative Pac I digests of one positively identified recombinant for each of the three shuttle vectors is shown in Figure 3.

Fig.3

High-Titer Virus Production and High-Level Gene Expression

Once recombinant adenoviral plasmid clones were identified, they were digested with Pac I and transfected into HEK293 cells using Stratagenes MBS Mammalian Transfection Kit, with modifications described by Pear and colleagues.4 HEK293 cells are human embryonic kidney cells that have been transformed by sheared Ad5 DNA.5 The cells contain and express the transforming genes of Ad5, including E1, which complements the deleted E1 ge
'"/>

Source:


Page: All 1 2 3 4 5 6

Related biology technology :

1. New Kit Generates Exceptionally Pure Total RNA for RT-PCR
2. Using XL1-Red Mutator Strain to Generate Esterase Variants
3. Fractionating DNA Fragments Generated by Differential Display PCR
4. Generate High-Quality, Directional Plasmid cDNA Libraries
5. High-Throughput System Generates Ultra-Pure PCR Products Suitable for All Applications
6. Inducing RNAi with siRNA Cocktails Generated by RNase III
7. Biolistic Gene Transfer to Generate Transgenic Schistosomes, Rev A
8. Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
9. Epitope-Tagging Vectors for Functional Analysis in Yeast
10. New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence Humanized Renilla GFP Reporter
11. Efficiently Insert Unique Restriction Sites into Plasmid Vectors
Post Your Comments:
(Date:8/27/2014)... , Aug. 27, 2014 ... the "Global Lactic Acid  (Biodegradable Polymer, Food ... Trends & Forecasts to 2019" report to ... simplest hydroxyl-carboxylic acid with an asymmetrical carbon atom. ... the biodegradable polymer, food and beverage, personal care ...
(Date:8/27/2014)... -- Reportbuyer.com has added a new market research ... Rational View https://www.reportbuyer.com/product/1595336/The-Graphene-Opportunity---A-Rational-View.html ... similar amount of hype currently surrounds graphene, with ... water treatment, but is graphene the only solution? ... Opportunity Report, authored by Tim Harper ...
(Date:8/27/2014)... Aug. 27, 2014 Tris Pharma, a specialty ... today announced that it has been selected as an ... Association,s 2014 Awards for Excellence competition in ... independent panel of judges from SCORE (Counselors to America,s ... will be honored at a gala event to be ...
(Date:8/27/2014)... August 27, 2014 Remcom announces an ... with new features for the biomedical industry. The ... Biological Thermal Sensor, exclusive to the Bio-Pro version of ... of XFdtd, developed to calculate the biological effects of ... been added to all versions to expand engineers’ post-processing ...
Breaking Biology Technology:Global Lactic Acid (Biodegradable Polymer, Food & Beverage, Personal Care & Pharmaceutical) Market - Trends & Forecasts to 2019 2The Graphene Opportunity - A Rational View 2The Graphene Opportunity - A Rational View 3The Graphene Opportunity - A Rational View 4Tris Pharma Selected as an award winner in the New Jersey Business & Industry Association's 2014 Awards for Excellence 2Remcom Announces Enhanced Biological Thermal Sensor and Other Biomedical Features in XFdtd EM Simulation Software Update 2Remcom Announces Enhanced Biological Thermal Sensor and Other Biomedical Features in XFdtd EM Simulation Software Update 3
... The Board of,Directors, meeting today under the chairmanship of Albert Saporta, approved,STALLERGENES, half year consolidated financial statements:, (EUR millions) ... ... % ... change, Sales ...
... NLTX ), today announced that Peter Strumph, Chief ... the Biotech Industry,conference on Thursday, September 4, at 1:30 ... City. Mr. Strumph will provide a company,overview and update ... The presentation will be webcast live and can be ...
... Aug. 27 Poniard,Pharmaceuticals, Inc. (Nasdaq: PARD ... that Jerry McMahon, Ph.D., chairman and CEO,will participate ... he,will provide a corporate overview, including a discussion ... lead product candidate., -- The Thomas Weisel ...
Cached Biology Technology:Stallergenes: First Half Year 2008: A 19% Increase in Operating Profit and Net Profit 2Stallergenes: First Half Year 2008: A 19% Increase in Operating Profit and Net Profit 3Nile Therapeutics to Present at BioCentury's NewsMakers in the Biotech Industry Conference 2Poniard Pharmaceuticals Announces Upcoming Conference Participation 2
(Date:8/27/2014)... America,s megafauna mastodons, short-faced bears, giant ground sloths, ... close to 13,000 years ago at the end of ... has long been debated by scientists who, until recently, ... of scientists, including UC Santa Barbara,s James Kennett, professor ... a comet collision with Earth played a major role ...
(Date:8/27/2014)... About 400 million years ago a group of fish began ... birds, and mammals. But just how these ancient fish used ... what evolutionary processes were at play remain scientific mysteries. ... Nature , turned to a living fish, called Polypterus ... first attempted to walk out of the water. Polypterus ...
(Date:8/27/2014)... invasive grass species frequently found in forests has created ... on American toads, a new University of Georgia study ... to the U.S. in the early 1900s, is one ... to more than a dozen states in the past ... and in forests, it can survive in widely diverse ...
Breaking Biology News(10 mins):Nanodiamonds are forever 2Nanodiamonds are forever 3Walking fish reveal how our ancestors evolved onto land 2More wolf spiders feasting on American toads due to invasive grass, UGA study shows 2More wolf spiders feasting on American toads due to invasive grass, UGA study shows 3
... in dogs in western and central Africa comes from ... years ago, probably by European colonialists. In the current ... team of scientists from Africa, USA and France report ... at country level and that there is only limited ...
... supports a modest beneficial effect of anakinra for rheumatoid ... infections and its use with other biologic medications. ... many as one in 100 people worldwide. It affects ... one of a new breed of arthritis drugs licensed ...
... operations necessary to treat stroke victims, confront hardened arteries ... be made safer as researchers from the Micro/Nanophysics Research ... to the design of micro-motors small enough to be ... research paper, published today, Tuesday, 20 January, in IOP ...
Cached Biology News:Hope for a rabies eradication strategy in Africa 2Anakinra for rheumatoid arthritis: A modest benefit with some risk 2Microbot motors fit to swim human arteries 2
... RNaseOUT Recombinant Ribonuclease Inhibitor is ... pancreatic-type ribonucleases such as RNase ... RNaseOUT Recombinant Ribonuclease Inhibitor is ... molecular weight ~52 kDa. RNaseOUT ...
... Natural and Recombinant RNasin Ribonuclease Inhibitors ... the inhibition of eukaryotic RNases of the ... its inhibitory effect by noncovalently binding to ... value for the binding of RNasin Ribonuclease ...
UGT1A7 (E-15)...
... Immunogen: synthetic peptide corresponding to the ... dopamine receptor, conjugated to KLH. The ... rat and mouse gene. Physical form: ... containing 0.01% sodium azide. Titertest method: ...
Biology Products: