Navigation Links
Mammalian Expression Vector for Efficient Cloning of PCR Fragments

CHO cells were harvested, luciferase activity was quantitated (figure 2) and the protein of the correct molecular weight was detected by Western blot analysis (data not shown). Luciferase activity and expression from the pCMV-Script vector were equivalent to that from the pcDNA 3.1 vector.

The pCMV-Script vector was stably transfected into CHO cells using Stratagenes Mammalian Transfection Kit. In separate experiments, the pCMV-Script vector and the pCMV-Script vector containing the luciferase gene were transfected into CHO cells, and G418 selection was used to obtain stable cell lines. Luciferase expression from stable cells containing the pCMV-Script vector with the luciferase gene insert was compared to cells containing the pCMV-Script vector with no insert. Detected luciferase activity was 1000-fold over background, measured by a standard luciferase assay (data not shown).

Conclusions

Stratagenes new pCMV-Script cloning kit offers high-efficiency cloning of blunt-ended PCR fragments for expression in mammalian cells. The pCMV-Script vector features (1) the CMV promoter for expression in a variety of cell lines, (2) the kanamycin- and neomycin-resistance gene for selection in either prokaryotic or eukaryotic systems and (3) ligation of insert to the pCMV-Script vector in the presence T4 DNA ligase and Srf I to eliminate background of recircularized vector. The pCMV-Script vector can be used to clone inserts that have been generated by any PCR enzyme. We have tested this vector and demonstrated high-level expression in mammalian cells.

REFERENCES

  1. Bauer, J., et al. (1992) Strategies 5: 62-65.

  2. Sanchez, T., Zheng, C.-F., and Bauer, J. (1996) Strategies 9: 44-46.

  3. Costa, G., and Weiner, M. (1994) Strategies 7: 47-48.


'"/>

Source:


Page: All 1 2 3 4

Related biology technology :

1. New Mammalian Two-Hybrid System Detects Protein-Protein Interactions
2. Highest Transfection Efficiency of an Endotoxin-Sensitive Mammalian Cell Line
3. An Epitope Tagging Vector for Gene Expression in Mammalian Cells
4. Versatile Vectors for Ponasterone A- Inducible Control of Gene Expression in Mammalian Cells
5. New Mammalian Expression Vectors Employ Stable, High-Level Fluorescence Humanized Renilla GFP Reporter
6. A New Lambda Vector for Mammalian Expression
7. Mammalian and Bacterial Expression in One Vector
8. Mammalian Expression Vectors with Interchangeable Selectable Markers
9. Low-Toxicity, Lipid-Mediated Transfection of Mammalian Cells
10. Mycoplasma Contamination Reduces the Effect of Lipid-Mediated Transfection of Mammalian Cells
11. Reagents for Up and Down-Regulation of miRNA Activity in Mammalian Cells
Post Your Comments:
(Date:12/17/2014)... (PRWEB) December 17, 2014 Gene ... that it has entered into a technology access ... (ADM) to apply DNA2.0’s proprietary protein engineering technology, ... process. , “We are extremely excited that the ... platform. This proprietary bioengineering technology has now ...
(Date:12/17/2014)... SOUTH SAN FRANCISCO, Calif. , Dec. 17, ... positive results of a Phase 2 study evaluating ... for the treatment of patients with severe, chronic ... the current standard of care, including topical steroids ... endpoint was percent change in Visual Analog Scale ...
(Date:12/15/2014)...  GlassesOff Inc. (OTCBB: GLSO) announced today the appointment ... director of the Company,s Board of Directors. ... its CEO until its acquisition by Stanley Black ... Recognized as the inventor of the first Wi-Fi-based Active ... -based RFID solutions focused on improving operational efficiency, safety ...
(Date:12/13/2014)... (PRWEB) December 12, 2014 Clarassance, a ... announced its new name: Therabron Therapeutics , Inc. ... and bronchioles (a type of structure in the lungs ... company’s mission to develop novel protein therapeutics for the ... directors decided to change the name to mark the ...
Breaking Biology Technology:ADM and DNA2.0 Enter Into Protein Engineering Technology Access and Service Agreement 2Velocity Pharmaceutical Development, LLC and Tigercat Pharma, Inc. Announce Phase 2 Results for VPD-737 in Patients With Chronic Pruritus 2Velocity Pharmaceutical Development, LLC and Tigercat Pharma, Inc. Announce Phase 2 Results for VPD-737 in Patients With Chronic Pruritus 3Velocity Pharmaceutical Development, LLC and Tigercat Pharma, Inc. Announce Phase 2 Results for VPD-737 in Patients With Chronic Pruritus 4AeroScout Founder Joins GlassesOff's Board of Directors 2AeroScout Founder Joins GlassesOff's Board of Directors 3Maryland-based Biotech Company's Path Forward in Treating Respiratory Diseases Sparks Name Change 2
... Morphotek®, Inc., a subsidiary of Eisai Inc., announced today that ... Cancer Care , a national support services organization for people ... educational programs for people facing lung and ovarian cancers.  The ... their families become aware of options available to help treat ...
... graphite particles 1000 times smaller than the width of a ... boost the efficiencyand profitabilityof solar power plants. Photovoltaic (PV) ... but they,re not necessarily the best solar power solution. "The ... only a fraction of the sunlight that hits them, and ...
... PARIS, April 5, 2011 Cellectis (Alternext: ALCLS), the genome ... signed an agreement with the Caisse des Depots (a French ... in two steps, reserved in equal parts to Cellectis and ... later than January 31, 2012, and is subject to the ...
Cached Biology Technology:Morphotek®, Inc. Receives Beacon Award From CancerCare 2Morphotek®, Inc. Receives Beacon Award From CancerCare 3Nanoparticles improve solar collection efficiency 2Cellectis and Caisse des Depots Sign Agreement on a Project for the Industrial Development of iPS Cells 2Cellectis and Caisse des Depots Sign Agreement on a Project for the Industrial Development of iPS Cells 3
(Date:12/17/2014)... NEW YORK , Dec. 15, 2014 /PRNewswire-USNewswire/ ... a Good Clinical Practices (GCP) audit to confirm ... Administration (FDA) regulations. This accomplishment enables HITLAB to ... trials utilizing the highest principles for patient safety ... to improve global healthcare access, quality, and delivery ...
(Date:12/11/2014)... 2014 Research and Markets ... ( http://www.researchandmarkets.com/research/twhjll/biometrics_market ) has announced the addition of ... 2015-2019"  report to their offering. ... advances in technology. With continuous advances in technology, ... the latest standard that meets the needs and ...
(Date:12/10/2014)...  Valencell, a leader in performance biometric data sensor ... from its licensees for highly accurate, clinically validated biometric ... coming from fitness and health sectors, but first responders/military ... is only as useful as the biometric data it ... in long-term mass consumer adoption of wearable products," said ...
Breaking Biology News(10 mins):HITLAB Completes GCP Audit for FDA Smart Device & App Compliance 2Biometrics Market in the APAC Region 2015-2019: Key Vendors are 3M Cogent, NEC, Safran and Suprema 2Wearable Technology Products Demand Highly Accurate Biometric Technology 2
... GAINESVILLE, Fla. New University of Florida research puts to ... the last glacial period. It turns out it ended up ... The findings have implications for modern-day global warming, said ... of the paper, which is published in this week,s journal ...
... team of researchers based at the Institute of Vertebrate ... professor at Washington University in St. Louis, has discovered ... anthropologists perceptions of the emergence of modern humans in ... published Oct. 25 in the online early edition of ...
... HOUSTON, Oct. 25, 2010 Some unexpected effects ... and reverse blindness have been uncovered by a University ... Donald A. Fox, a professor of vision sciences ... findings in a paper titled "Low-Level Gestational Lead Exposure ...
Cached Biology News:UF research gives clues about carbon dioxide patterns at end of Ice Age 2UF research gives clues about carbon dioxide patterns at end of Ice Age 3Unexpected findings of lead exposure may lead to treating blindness 2Unexpected findings of lead exposure may lead to treating blindness 3