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. 2014 Mar 21;9(3):e92377. doi: 10.1371/journal.pone.0092377

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© 2014 Feenstra et al

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) ΔD(S2del) virus with the GFP sequence inserted (ΔD(S2delGFP)) was generated. GFP expression was obvious during several successive virus passages in BSR cells, as shown for passage 6 and 7 (p6, p7). GFP expression was less obvious after subsequent passages, as shown for passages 8 and 9 (p8, p9). (B) Genetic stability of Seg-10 of ΔD(S2delGPF) during ten passages was studied by RT-PCR amplification of Seg-10. The original Seg-10 of ΔD(S2delGFP) mutant virus was identified (.), but in subsequent passages additional smaller amplicons became more prominent (*). The middle small band has a deletion in the GFP sequence, the smallest amplicon has a larger deletion in the GFP sequence, and in the largest of the small amplicons, the Seg-2 insertion is also deleted, but a Seg-6 sequence is inserted instead.