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

ne from the pAdEasy-1 vector for assembly of replication-defective adenovirus. To confirm the presence of infectious virus, primary viral stocks were subsequently used to transduce HEK293 cells. Three days after transduction, cells were analyzed for expression of their respective reporter genes. Cells transduced with AdCMV-GFP and Ad-CG (both harboring a GFP reporter gene) were visualized by fluorescence microscopy. Cells transduced with AdCMV-LacZ were stained with X-gal using Stratagenes In Situ b-Galactosidase Staining Kit. From representative photographs of HEK293 cells transduced with the test vectors (Figure 4), the positive identification of the reporter gene expression in transduced cells indicates that infectious adenovirus were produced using the AdEasy vector system.

Fig.4

Conclusions

Stratagenes AdEasy vector system simplifies and speeds up the process of generating a recombinant adenovirus, compared to traditional methods of preparation. With the AdEasy systems unique two-step process, a gene of interest, or expression cassette, is subcloned into a shuttle vector, then transferred into the adenoviral genome by homologous recombination in E. coli. This method eliminates the need for laborious plaque isolations, since the resulting adenoviral plasmid can be directly transfected into HEK293 cells to prepare adenovirus. With the AdEasy system, adenoviral-mediated gene transfer and expression are realized efficiently and conveniently.

REFERENCES
  1. Benihoud, K., Yeh, P., and Perricaudet, M. (1999) Curr. Opinion in Biotech. 10: 440-447.

  2. Berkner, K. (1988) Biotechniques 6: 616-625.

  3. He, T., et al. (1998) Proc. Natl Acad Sci USA 95: 2509-251
    '"/>



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:4/21/2015)... (PRWEB) April 21, 2015 Staying true ... to craft a digital product that makes the world ... with Global Biological Standards Institute (GBSI) to redesign their ... clean, and most importantly, strategy-based. , "Biomedical research is ... - much less to promote it in a narrative ...
(Date:4/21/2015)... , April 21, 2015 ... and development, custom automation, and contract manufacturing, and  ... accessible flow cytometry and cell sorting technology for ... develop a novel flow cytometry platform. ... NanoCellect Biomedical has developed a microchip-based cell sorting ...
(Date:4/21/2015)... , April 21, 2015  Tute Genomics, a leading ... of Josh Forsythe as VP of Marketing. Mr. ... witnessed a significant expansion in its commercial operations over the ... " Josh Forsythe is a tremendous addition ... MD MBA, CEO of Tute Genomics. "Josh,s vast experience commercializing ...
(Date:4/21/2015)... and RESEARCH TRIANGLE PARK, N.C. , ... UTHR ) announced today that it will release ... on Tuesday, April 28, 2015. United Therapeutics ... 2015, at 9:00 a.m. Eastern Time.  The teleconference is ... A rebroadcast of the teleconference will be available for ...
Breaking Biology Technology:Visceral Tells GBSI’s Story Within 30 Seconds of Attention 2Visceral Tells GBSI’s Story Within 30 Seconds of Attention 3NanoCellect And Invetech Announce Collaboration To Develop Novel Flow Cytometry Platform For Cell Sorting And Analysis 2NanoCellect And Invetech Announce Collaboration To Develop Novel Flow Cytometry Platform For Cell Sorting And Analysis 3Tute Genomics Continues Expansion of Genomic Medicine Commercial Operations; Appoints Josh Forsythe as New VP of Marketing 2Tute Genomics Continues Expansion of Genomic Medicine Commercial Operations; Appoints Josh Forsythe as New VP of Marketing 3Tute Genomics Continues Expansion of Genomic Medicine Commercial Operations; Appoints Josh Forsythe as New VP of Marketing 4
... New Haven, Conn.A team led by Yale University scientists ... different cells in the blood using magnetizable liquids. The ... 7 in the online edition of the Proceedings ... improve the speed and sensitivity of tests used to ...
... quickly produce ultra-lightweight, bendable batteries and supercapacitors in ... a sheet of paper with ink made of ... conductive storage device, said Yi Cui, assistant professor ... needs a low-cost, high-performance energy storage device, such ...
... , QUINCY, Mass., ... International), a leading biotech developer and manufacturer of renewable biochemicals, ... to receive up to $50 million for its planned biobased ... integrated biorefinery projects that won funding or loan guarantees, Myriant ...
Cached Biology Technology:New technology could boost disease detection tests' speed and sensitivity 2Dip ordinary paper into ink infused with nanotubes and nanowires to create an instant battery 2Myriant Technologies LLC Selected for $50 Million Award for Succinic Acid Biorefinery Project 2
(Date:3/31/2015)... 2015   Guidepoint , a leading global research ... Pain Management TRACKER, part of the Guidepoint TRACKER ... the medical device and therapeutics markets. The Post-Surgical Pain ... volumes, market share, and adoption rates across three major ... surgery. The Post-Surgical Pain Management TRACKER will ...
(Date:3/26/2015)... The Granite Club, Canada,s ... announced it has implemented a state-of-the-art access and security ... and staff, while restricting access to authorized users only. ... IMID Access system because it has the perfect combination ... in addition to unparalleled security," said James DiRenzo ...
(Date:3/24/2015)... 24, 2015 Research and Markets ( http://www.researchandmarkets.com/research/kvdtcw/face_and_voice ) ... - Global Strategic Business Report" report to their offering. ... Biometrics in US$ Thousands by the following Segments: Face Biometrics, and ... Canada , Japan , ... Middle East & Africa , and ...
Breaking Biology News(10 mins):Guidepoint Launches Post-Surgical Pain Management TRACKER 2Guidepoint Launches Post-Surgical Pain Management TRACKER 3Granite Club First Private Club in North America to Implement FST Biometrics Secure Access Solution, as Part of Multi-million Dollar Expansion 2Face and Voice Biometrics - Global Strategic Business Report 2015: Face Recognition Gains Prominence in Immigration Control 2Face and Voice Biometrics - Global Strategic Business Report 2015: Face Recognition Gains Prominence in Immigration Control 3Face and Voice Biometrics - Global Strategic Business Report 2015: Face Recognition Gains Prominence in Immigration Control 4Face and Voice Biometrics - Global Strategic Business Report 2015: Face Recognition Gains Prominence in Immigration Control 5Face and Voice Biometrics - Global Strategic Business Report 2015: Face Recognition Gains Prominence in Immigration Control 6
... The ecological value of coastal mangrove forests in Mexico has ... first time, researchers have used a wide-ranging compilation of fisheries ... economic price tag on that value. Scientists at ... the new online edition of the Proceedings of the ...
... spread well beyond the continental shelf and drive carbon capture ... multi-year study. The finding does not change estimates of ... large role of tropical oceans and major rivers. The ... carbon from the respiration of ocean life. A 2007 study ...
... engineers have developed a new imaging system that allows ... mutation. The work, which could help scientists understand ... have been able to pinpoint the number and location ... In this case, the researchers worked with mouse pancreatic ...
Cached Biology News:Scripps study sets high economic value on threatened Mexican mangroves 2Scripps study sets high economic value on threatened Mexican mangroves 3Amazon powers tropical ocean's carbon sink 2MIT researchers offer glimpse of rare mutant cells 2
Rabbit polyclonal to XTP4 ( Abpromise for all tested applications). entrezGeneID: 84299 SwissProtID: Q9BRT3...
Chicken polyclonal to QPRT ( Abpromise for all tested applications). Antigen: Synthetic peptide, corresponding to Internal sequence amino acids 123-237 of Human QPRT Entrez Gene ID: 23475 ...
...
Antibody Diluent for IHC 125 ml...
Biology Products: