Transfection of REV-A and HTLV-1 bicistronic reporters yield heterogeneous transcript population
Taken from "RNA helicase A interacts with divergent lymphotropic retroviruses and promotes translation of human T-cell leukemia virus type 1"
Nucleic Acids Research 2007;35(8):2629-2642.
Published online 10 Apr 2007
© 2007 The Author(s)Diagram of CMV IE-driven bicistronic reporter gene that encodes renilla luciferase (R-luc) in the first cistron, firefly luciferase (F-luc) in the second cistron and indicated intercistronic regions. Representative northern blot with F-luc DNA probe verifying expression of the expected ∼3.6 kb transcript and aberrant transcripts marked with an asterisk. Four independent transfection assays were performed in HeLa cells and total cellular proteins were harvested at 48 h post-transfection and subjected to dual Luciferase assay. Box plot analysis summarizes the range of F-Luc activity with internal line indicating mean and extending lines connecting extreme values. Scatterplot analysis of the four replicate experiments demonstrated that pPolio and pREV-A and pHTLV-1 segregated separately from the pNoIRESfs plasmid. Natural log transformation was used to adjust non-normality and unequal variances. Northern blot with F-luc DNA probe indicates aberrant transcripts are also present in transfected D17 cells.