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. 2019 Apr 26;9:126. doi: 10.3389/fcimb.2019.00126

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Copyright © 2019 Liu, Lei and Kadowaki.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Comparison of disrupting LpMT by CRISPR/Cas9-induced HDR and homologous recombination. (A) Wild type (WT) and truncated (Truncation) alleles of LpMT are shown as in Figure 5A. The positions of primers to detect LpMT truncated allele are shown with the expected size of PCR product. (B) Ten drug (blasticidin, G418, and hygromycin) resistant clones (1–10) isolated by CRISPR/Cas9-induced HDR together with wild type L. passim were analyzed by genomic PCR to detect WT (851 bp amplicon) and Truncation (803 bp amplicon) alleles of LpMT. The positions of 500 and 1,000 bp bands in molecular weight marker (MW) are shown at the right. (C) Ten hygromycin resistant clones (1–10) isolated by homologous recombination were analyzed as in (B).