Figure 4
From: Characterization of a synthetic human LINE-1 retrotransposon ORFeus-Hs
Analysis of ORF1 protein expression and relative ORF2 RT activity. (A) The same constructs as in Figure 2 were analyzed. The vectors used were: pLD223, pWA172, pWA170, pWA176, pWA163, pWA174, pLD224, pLD227, pLD225, pWA165. Top, protein expression of ORF1. Bottom, protein expression of the tubulin loading control. (B) L1 element amplification protocol (LEAP): assay was performed using ribonucleoproteins (RNPs) prepared from cells transfected with pWA174 (L1RP) and pWA165 (ORFeus-Hs). The numbers 0, 0.1, 0.2, 0.5, 1, 5 and 10 indicate the amount (μg) of total protein of the RNP prep added to each LEAP reaction. The arrows indicate the mobility of LEAP PCR product. SuperScript III represents a positive control in which 100 U of SuperScript III reverse transcriptase (Invitrogen, Carlsbad, CA, USA) was added to the LEAP reaction.