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Plasmids 101: Viral Vector Elements

Posted by Marcy Patrick on Jul 17, 2014 3:09:00 PM

The use of viral vectors in research is beneficial for a number of reasons, including but not limited to: helping to get difficult-to-deliver DNA into mammalian cells, increasing the efficiency of gene transduction, allowing for control over which cells are infected through viral pseudotyping, and ease of vector cloning and modification. At the most basic level, viral vectors consist of a viral genome that has been adapted into a plasmid-based technology and modified for safety through the removal of many essential genes and the separation of the viral components. Read on for a brief description of the viruses used to make these vectors as well as a table defining the major elements found within the plasmids comprising the viral vector systems.

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Topics: Plasmid Elements, Plasmids 101, Viral Vectors

Plasmids 101: Luciferase

Posted by Jason Niehaus on Jun 24, 2014 11:59:00 AM

Luciferases are a class of enzymes capable of catalyzing chemical reactions in living organisms resulting in the emission of photons. The most familiar bioluminescent organism for most people is the firefly (Photinus pyralis) and perhaps not surprisingly it is also the most commonly used bioluminescent reporter. This beetle emits a yellow-green light with a peak emission at 560nm. Shortly after the initial article describing the cloning of firefly luciferase was published in 1985, several studies utilized luciferase as a genetic reporter in plant and mammalian cells. Luciferase assays have since become a gold standard in gene expression analysis and a luciferase gene (one of many available to choose from) is now a common feature in reporter plasmids. 

Learn How Luciferase Can Be Used In Concert with Fluorecent Proteins in Nano Lanterns

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Topics: Plasmid Technology, Plasmids 101

Plasmids 101: Green Fluorescent Protein (GFP)

Posted by Marcy Patrick on May 15, 2014 11:33:00 AM

Bioluminescence and fluorescence from proteins such as Green Fluorescent Protein (GFP) has likely existed in creatures such as jellyfish for millions of years; however, it took until the 1960s for scientists to begin to study GFP and deduce its biochemical properties. Now GFP and its fluorescent derivatives are a staple in the lab. GFP is used in research across a vast array of biological disciplines and scientists employ GFP for a wide number of functions, including: tagging genes for elucidating their expression or localization profiles, acting as a biosensor or cell marker, studying protein-protein interactions, visualizing promoter activity, and much more.

Read on to learn more about GFP, how scientists have evolved this versatile protein to suit their experimental needs, and some of the common applications in the lab.

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Topics: Plasmid Elements, Plasmids 101, Fluorescent Proteins

Plasmids 101: The Promoter Region – Let's Go!

Posted by Kendall Morgan on Apr 3, 2014 4:05:00 PM

Thus far in our Plasmids 101 series we've worked our way through the plasmid map: antibiotic resistance, origin of replication, and so on. Up to this point we can replicate our plasmid and make sure cells maintain it; the next step is getting the plasmid to express our gene of interest. Enter the promoter-- the element responsible for initiating the transcription of your insert into RNA.

In practice, the term "promoter" describes the combination of the promoter (RNA polymerase binding site) and operators (response elements). Promoters are about 100 to 1000 base pairs long and found upstream of their target genes. The sequence of the promoter region controls the binding of the RNA polymerase and transcription factors, therefore promoters play a large role in determining where and when your gene of interest will be expressed. 

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Topics: Plasmid Elements, Plasmids 101

Plasmids 101: Mammalian Vectors

Posted by Marcy Patrick on Mar 25, 2014 11:15:00 AM

Although plasmids do not naturally exist in mammals, scientists can still reap the benefits of plasmid-based research using synthetic vectors and cultured mammalian cells. Of course, these mammalian vectors must be compatible with the cell type they are tranfected into – a bacterial origin of replication (ORI) will not allow for plasmid replication in mammalian cells, for example, and a toxin that kills bacteria may not have any discernable effect on mammalian cells. In this blog post we will discuss how mammalian plasmids differ from their bacterial counterparts, including how replication occurs and whether selection is necessary for transfected cells.

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Topics: Plasmid How To, Plasmid Technology, Plasmids 101

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