Figure 4 of
Sullivan, Mol Vis 2002;
Figure 4. Second generation hRzs Cleave long WT and C187Y opsin mRNAs during co-synthesis/cleavage
Linearized plasmid templates containing opsin or hRz cDNAs downstream of the T7 promoter were mixed in a cDNA template molar ratio of 1:6 ([E]:[S]) and in vitro co-transcription/ cleavage reactions were conducted for 1 h at 37 °C. The reactivities of anti-V230 Rz2A and Rz2B on both WT and C187Y full length mRNAs (S) are shown. Cleavage products (Ps) of expected sizes (P1=782 nt and P2=1056 nt) were generated with each hRz reacting with each mRNA target. C187Y and WT mRNAs cut with Rz2A are in lanes 1 and 2, respectively, and the same targets cut with Rz2B are in lanes 3 and 4. C187Y and WT synthesized without hRz plasmid are in lanes 5 and 6, respectively. Lane 7 is an RNA ladder. hRz RNAs (Rz2A 55 nt, Rz2B 61 nt) are not seen in this gel image. The 1.2 kB band seen on all lanes (except 5) is nonspecific and may reflect an incomplete transcript generated by T7 RNA polymerase stalling.