Figure 1
From: RNA mutagenesis yields highly diverse mRNA libraries for in vitroprotein evolution
Qβ replicase mutagenesis/ribosome display vector and typical Qβ replicase amplification reaction. (A): Schematic representation of plasmid pEGX253. Base plasmid pEGX216 (elements depicted with underline) comprised of a MCS site inserted into a modified RQ 135-1(-) sequence which was then used to add the required elements to construct pEGX253. Target gene sequence (12Y-2) was cloned into the NcoI and NotI restriction sites. (B): Product from a typical Qβ replicase amplification of RGS mRNA. Lane 1 represents the amount of RGS single-stranded mRNA (-) template added to the Qβ replicase reaction. Lane 2 shows the Qβ replicase reaction product following amplification for 2 hrs at 37°C. Note that Qβ replicase amplifies both (-) and the newly generated (+) strands of the RNA eventually leading to a dsRNA product.