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. 2014 Jul 23;5:369. doi: 10.3389/fmicb.2014.00369

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Copyright © 2014 Karas, Wise, Sun, Venter, Glass, Hutchison, Smith and Suzuki.

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) or licensor 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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Colony phenotypes associated with nusG deletion and rescue. (A–C) Individual colonies were photographed 72 h after plating on agar medium from sparse fields where colony growth would be unaffected by neighboring colonies. Colonies represent (A) JCVI-syn1.0, (B) JCVI-syn1.0 ΔL, and (C) complemented clone ML1. (D) represents a field of colonies from the second culture in Figure 4, showing the initial emergence of the faster growing colonies among the more prevalent smaller colonies within the population derived from transformation of JCVI-syn1.0 ΔL using the marker-less approach. Centers of colonies display the typical mycoplasma “fried-egg” characteristic. Blue color is from expression of lacZ in medium containing X-gal (smaller colonies had not yet developed color at the time of assay). Bars represent 500 μm.