To verify the promoter activity of these genomic regions, we designed a dual reporter vector in which mCherry and EGFP were cloned in sense and anti-sense orientation to genomic elements with predicted promoter activity (Fig. 4e).
However, a second antibiotic is not required for expression of the dual vector in standard BL21 competent cells.
A dual-vector approach, in which the Factor VIII heavy chain and light chain are separately encoded by two different vector genomes, is another mechanism by which the large transgene can be accommodated (Mah et al, 2003).
RT sequences coding for the p66 and p51 subunits of BH10 were cloned in the pRT dual vector, which is derived from pCDF-2 with LIC duet minimal adaptor (Novagen), using restriction sites PpuMI and SacI for the p51 subunit, and SacII and AvrII for the p66 subunit.
To confirm that miRNA-1 is able to functionally bind to 3′ UTR of Clcn3 and modulate it's expression, we constructed a luciferase reporter vector in which the 3′UTR of Clcn3 mRNA was fused to the luciferase coding sequence under pGK promoter in dual luciferase vector (pmiRGLO vector).
A symplectic system is established directly by introducing dual vectors in terms of the symplectic eigenfunctions.
Instead, a dual rAAV vector system in which the TRE-TRAIL and rtTA were constructed into two rAAV vectors respectively provided a similar intensity in transgene expression and much more strict controllability compared with the single vector system.
Thus, we chose to construct a new set of piggyBac vectors in which pairs of heterologous genes could be placed under the control of the dual ie1-hr5-ie1 control element.
Combinatory effects of replication-competent Ad and forced expression of p53 can be examined either with dual transduction using 2 kinds of Ad vectors or with a single vector system in which the p53 gene was integrated into the replication-competent Ad vector.
(Supplemental Figure S2) Studies here demonstrate the limitations of dual vector delivery in difficult fibroblast transfection conditions.
Protein ligation by the new split inteins was tested in vivo using the dual vector system previously developed in our group [8].
