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. 2016 Apr 15;30(8):909–917. doi: 10.1101/gad.278432.116

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© 2016 Carmona et al.; Published by Cold Spring Harbor Laboratory Press

This article is distributed exclusively by Cold Spring Harbor Laboratory Press for the first six months after the full-issue publication date (see http://genesdev.cshlp.org/site/misc/terms.xhtml). After six months, it is available under a Creative Commons License (Attribution-NonCommercial 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/.

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Figure 3. — Recombination mediated by HzTransib and purple sea urchin RAG1-like. (A,B) Quantitation of recombination in RAG1^−/− 3T3 fibroblast lines. Recombination efficiency is plotted as in Figure 1B. Note that levels of recombination by RAG1 alone in this RAG1^−/− line were higher than in wild-type and RAG2^−/− lines. (Mock) Empty vector control; (HZ) Hztransib; (SpR1L) spRAG1L; (HZm) active site mutant of Hztransib; (SpR1Lm) active site mutants of spRAG1L. (C) Representative PCR assays for coding or signal joints in genomic DNA from MPA-resistant cells expressing the indicated proteins. Each lane is an independent clone. (Plasmid) Reaction spiked with positive control template DNA; (HZ) Hztransib; (SpR1L) spRAG1L. (D) GST pull-down to detect interactions between the indicated MBP- or GST-tagged proteins. (R1c) RAG1 core; (Hz) HZTransib; (mR2) mouse RAG2; (sR2) shark RAG2; (+) GST alone. (Top) Glutathione pull-down. (Middle and bottom) Starting material lysate. Arrows indicate the expected positions of MBP-RAG1 core and MBP-Hztransib proteins.