期刊
ARCHIVES OF VIROLOGY
卷 154, 期 5, 页码 853-856出版社
SPRINGER WIEN
DOI: 10.1007/s00705-009-0368-y
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- Polish Ministry of Science [N N 302 312 934]
For the first time, a full-length cDNA clone of the RNA genome of pepino mosaic virus (PepMV) was constructed. RNA was extracted from purified virions of isolate PepMV-Pa and used for cDNA synthesis. The full-length cDNA was produced as one 6.4-kb fragment representing the entire PepMV genome. This fragment was ligated into the pCR(A (R)) -XL-TOPOA (R) vector downstream of T7 RNA polymerase promoter, which was included in the 5' primer sequence used for RT-PCR. The PepMV-Pa RNA transcripts obtained were infectious in different host plants, causing symptoms indistinguishable from those of the wild-type isolate. The presence and authenticity of the progeny virus were verified by ELISA, RT-PCR and nucleotide sequencing.
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