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. 2019 Nov 7;9:16274. doi: 10.1038/s41598-019-52822-1

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Identification of the Bgl II restriction site in rCBPV RNA2. (a) A RT-PCR fragment flanking a Bgl II site, which was introduced in the rCBPV sequence as a genetic marker, was performed using the primers CPV33 and CPV34. Pupae transfected with recombinant CBPV (lane a2), infected with passaged rCBPV (a3, a4) or infected with wild type CBPV (lane a5 and a6) show an RT-PCR product of the expected size (546 bp). PBS injected pupae serve as negative controls with no visible amplicon (lane a1 and b1). (b) Digestion of the RT-PCR products with Bgl II yielded two distinct bands of 327 and 219 bp in the case of recombinant CBPV (lane b8-b10), compared to wtCBPV, where no cleaved PCR product occurred (lane b11 and b12).