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Transfection of REV-A and HTLV-1 bicistronic reporters yield heterogeneous transcript population

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posted on 31.12.2011, 07:18 by Cheryl Bolinger, Alper Yilmaz, Tiffiney Roberts Hartman, Melinda Butsch Kovacic, Soledad Fernandez, Jianxin Ye, Mary Forget, Patrick L. Green, Kathleen Boris-Lawrie

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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

PMCID:PMC1885656.

© 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.

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