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. 2010 Oct 28;6(10):e1001161. doi: 10.1371/journal.ppat.1001161

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Lye et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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Panel A. LUC RNAi reporter assays. pIR2SAT constructs expressing LUC alone (black boxes) or the LUC RNAi self reporter (LUC SR; white boxes) were introduced separately into L. braziliensis M2903, L. guyanensis M4147 (LRV1-4 virus-containing), or L. guyanensis M4147/pX63HYG (virus-free). SSU-integrated clonal lines were obtained and assayed for luciferase activity (n = 4 for M2903; n = 6 for L. guyanensis; the average and standard deviations are shown). The ratio of luciferase activities between the LUC SR and LUC expressing clones of each of the three lines are shown below the graph. Panel B. PCR confirmation of LRV1-4 virus status in parental and transfectant L. guyanensis M4147. PCR primers were LRV1-4 set 1 (lanes 3,5,7,9,11) or set 2 (lanes 2,4,6,8,10,12) (Table S1). RT-PCR reactions were performed with RNAs isolated from L. braziliensis M2903 (virus-free control; lanes 1,2), M4147 (obtained from two sources; lanes 3,4 and 5,6), M4147+LUC SR (lanes 7,8), M4147/pX63HYG (lanes 9,10), or M4147/pX63HYG + LUC SR (lanes 11,12). M, molecular size marker.