Team:Freiburg Bioware/BioBricks/vector plasmid



Vectorplasmid BioBricks

Vectorplasmid BioBricks allow it to reconstitute a vectorplasmid of your choice. Either the provided parts are assembled or an other Gene of Interest (GOI) can be integrated into an vectorplasmid. Beside the packaging capacity of 4.7 kbb, the possiblilities are virtually endless.
The Inverted Terminal Repeats (ITRs) are constructs form highly stable secondary structures that are recognized by the Rep proteins and function as a packiging signal. There for the ITRs have to flank the later vectorplasmid.
The Beta-globin and the human growth hormon polyadenylation sequence (hGH)are regulatory sequences that are ment to enhace the expression of the used GOI.

Create your own Vectorplasmid with the PrecursorBricks

For several applications users of the Virus Construction Kit will only want to insert their own GOI and not change the promoter and regulatory elements in a first experiment. For these applications we provide precursors of our vectorplasmid that reduce the required cloning on two steps.

Assembled Vectorplasmid - ready for transfection

For the development of retargeted viral vectors the vectorplasmid often functions as generator for different read-outs that can be measured in several assays. For this purpose it is often desired to use fully assembled vectorplasmids that are confirmed working and provieded in our Virus Construction Kit.
There for we assembled vectorplasmids with either fluorescent proteins like mCerulean(CFP), mVenus(YFP) and mCherry or produrg convertases like the cytosine deaminase or the tymindine kinase.
Detection of productive infections is possible by measuring the fluorescence of the fluorescent proteins or the number of killed cells where as the detection of the vectorplasmid is in any step of the infection possible by performing a qPCR reaction with primers for the CMV promoter.