pBIND 载体

The pBIND Vector is a high-copy plasmid in which the CMV immediate early promoter drives expression of a portion of the yeast GAL4 gene (amino acids 1–147) containing a DNA-binding domain. As with the pACT Vector, the DNAbinding domain sequence is flanked by a chimeric intron, a multiple cloning region, stop codons and an SV40 late polyadenylation region (Figures 4 and 5). The fusion gene region is flanked by T7 and T3 RNA polymerase promoters for the purpose of synthesizing sense and antisense RNA products, respectively. The Renilla luciferase gene on this vector is preceded by the SV40 early promoter and a growth hormone intron. Introns can increase protein expression through mRNA stability and nuclear to cytoplasmic transport effects (9–12). A synthetic polyadenylation sequence resides 3´ of the Renilla luciferase gene. The plasmid backbone contains an f1 origin of replication for the production of ssDNA and the β-lactamase gene (Ampr) for selection in E. coli.