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Highly Precise and Developmentally Programmed Genome Assembly in Paramecium Requires Ligase IV–Dependent End Joining

Figure 7

Absence of final junction products following IES excision in LIG4-silenced cells.

IESs #1 ( = 51A1835), 3 ( = 51A4404) and 6 ( = 51G4404) are described in the legend to Figure 6. Two additional IESs from surface antigen A gene are presented: 2 = 51A2591 (370 bp) and 4 = 51A4578 (883 bp). In each diagram, IESs are represented by a grey box and their flanking MAC sequences by a black line. PCR primers are drawn as arrowheads. A. Detection of IES excision junctions in the A gene in the macronuclear variant 51ΔA. Top: gel electrophoresis of PCR products around IESs. Bottom: representation of MAC development stages. In vegetative cells, only the MIC contributes to the PCR signal (IES+). When new MACs develop, de novo chromosomal junctions give an IES- PCR signal. At later stages, this signal disappears due to maternal epigenetic inheritance of the A deletion [13]. B. Detection of IES circle junctions by PCR in 51ΔA, using two internal divergent primers for each IES (same time-course as in A).

Figure 7