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This can be achieved by delivering a high concentration of vector in a minimal volume.
Cells were transiently transfected by electroporation with various concentration of vector coding for human V1a receptor and empty vector to a final amount of 10 íg.
Since the empty vector controls showed no evidence of neurodegeneration we concluded that expression of a protein from this high concentration of vector was sufficiently burdensome to be toxic to SN DA neurons.
At the lowest concentration of vector, KLF15 is mainly over-expressed in the nucleus fraction.
However, KLF15 expression induced a 50% decrease of luciferase activity, whatever the concentration of vector.
A certain concentration of vector mosquitoes is needed to cause a dengue epidemic.
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Interestingly, these results suggest that Ad49 can transduce vascular cells with greater efficiency than Ad35 and that lower concentrations of vector than have been previously used may be sufficient in vivo.
100 µl of sc-AAV5-smCBA-hGFP vector at a concentration of 4.5×1013 vector genomes/ml (produced as described by Zolotukhin, et al. [8]) was injected into the potential space.
An application of this approach to the study of vector-borne diseases would have to include the spatio-temporal correlations in the concentrations of vectors and infected individuals.
Briefly, putative enhancer fragments in the pGL3 promoter vector (Promega) were cotransfected into SW948 cells with 1/10 concentration of pGL4 Renilla vector.
This was followed by application of a biotinylated secondary antibody for 24 hours at a concentration of 1∶200 (Vector Labs, Burlingame, CA).
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