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. 2014 Jul 17;14:65. doi: 10.1186/1472-6750-14-65

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Copyright © 2014 Lerche and Hallmann; licensee BioMed Central Ltd.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

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Demonstration of co-transformation of heterologous genes. Paromomycin-resistant transformants were analyzed for the presence of the co-transformed, non-selectable DNA in the genome. PCRs were conducted using genomic DNA from transformants co-transformed with the pPsaD-GLuc (A), pHRLucP (B) or pHsp70A-GLuc (C) plasmids as template. The parental wild-type strain was analyzed as a control. Primers were specific for the heterologous gluc gene, and a 342-bp PCR fragment (Figure 3B-D) was expected in co-transformants. The rightmost lane shows a positive control using DNA of the co-transformed plasmid as the template. Lane M refers to the molecular weight marker. (D) Sequence obtained from the amplified and cloned gluc fragments. Primer positions, the stop codon (bold), and a BamHI restriction site (italics) are indicated.