Construction of the vectors pW-bc and pE3SP-bc

Copyright information:

Taken from "Development of a species-specific RNA polymerase I-based shRNA expression vector"

Nucleic Acids Research 2006;35(2):e10-e10.

Published online 07 Dec 2006

PMCID:PMC1802596.

© 2006 The Author(s).

Part of the pW ribosomal minigene was PCR amplified in two steps. First, two fragments were generated using primers P1 and P2, or P3 and P4, respectively. Second, both fragments were joined by PCR with primers P1 and P4. The restriction sites PstI and EcoRI, introduced through PCR primers P1 and P4, were used to insert the resulting fragment into the pW and pE3SP (pE) vector backbones. P, mouse Pol I promoter; Tx, transcription termination signal; bc, basic construct; E, enhancer.