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In one system, the Cre protein is expressed from the retrofitting plasmid itself in vivo.
The luciferase cassette is expressed efficiently from the retrofitting plasmids and from retrofitted BACs after transient transfection of B16F10 cells in tissue culture and after electroporation into muscles of BALB/c mice in vivo.
The conference will also feature tours and receptions to call attention to New York's green advances, from the retrofitting of the Empire State Building to construction of the High Line, a public park on the site of a former freight railway line.
However, prior to insertion of DNA onto a BAC at a loxP site, it is necessary to remove or inactivate any high-copy origins from the retrofitting DNA as such an origin would make the BAC multicopy.
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Alternatively the Cre can be expressed from a separate plasmid that is co-transfected with the retrofitting plasmid, and then lost once retrofitting has occurred.
The retrofitting plasmids give almost identical luciferase expression to the control pCMV-LUC and pCIKLux plasmids.
This ensured the simulation of a very challenging condition for the retrofitting from an energetic point of view (i.e., for moderate or low separation factor membranes: the higher the flux, the higher the process energy requirement).
The specific primers used for the retrofitting of each construct are listed in Table 4.
These are just some of the lists that the comic strip "XKCD" recently joked would result from retrofitting the twentieth century's most newsworthy events with modern, Internet-style headlines.
"6 TITANIC SURVIVORS WHO SHOULD HAVE DIED". "THESE 9 NAZI ATROCITIES WILL MAKE YOU LOSE FAITH IN HUMANITY". "5 INSANE PLANS FOR FEEDING WEST BERLIN YOU WON'T BELIEVE ARE REAL". These are just some of the lists that the comic strip "XKCD" recently joked would result from retrofitting the twentieth century's most newsworthy events with modern, Internet-style headlines.
Those steps run the gamut from retrofitting buildings for seismic resistance; strengthening governance practices on matters of zoning and building codes; helping Nepal develop a more robust disaster response management capacity; creating better advance preparations for search and rescue to save more lives; and raising public awareness to the steps individuals can take to prepare themselves better.
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Justyna Jupowicz-Kozak
CEO of Professional Science Editing for Scientists @ prosciediting.com