Potato virus Y (PVY) full-length cDNA has been found to be refractory to cloning in Escherichia coli cells. A full-length 9.7 kb PW cDNA was obtained by reverse transcription polymerase chain reaction (RT-PCR) from the RNA of PVY (tuber necrotic strain, PVYNTN). Double-stranded DNA fragments were used as primers (ds megaprimers), to include signals for transcription in vivo (a cauliflower mosaic virus 35S RNA promoter and a nopaline synthase terminator) in the final PCR product. Biolistic bombardment with a helium particle gun was used to inoculate the amplified product to detached tobacco leaves. Inoculation of tobacco plants with ground inoculated leaves followed by northern blot, ELISA and immune-electron microscopy demonstrated that the DNA was highly infectious with up to 90% of bombarded leaves containing the virus. This methodology will allow the use of reverse genetics in the study of PVY-plant interactions and will also be useful for obtaining infectious cDNA from other viruses with large RNA genomes.
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UNIV PARIS 07, INST JACQUES MONOD, INST MIXTE, CNRS, F-75251 PARIS 05, FRANCEUNIV PARIS 07, INST JACQUES MONOD, INST MIXTE, CNRS, F-75251 PARIS 05, FRANCE
BOYER, JC
HAENNI, AL
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UNIV PARIS 07, INST JACQUES MONOD, INST MIXTE, CNRS, F-75251 PARIS 05, FRANCEUNIV PARIS 07, INST JACQUES MONOD, INST MIXTE, CNRS, F-75251 PARIS 05, FRANCE
机构:
WASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USAWASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USA
CHENG, S
FOCKLER, C
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WASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USAWASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USA
FOCKLER, C
BARNES, WM
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WASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USAWASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USA
BARNES, WM
HIGUCHI, R
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WASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USAWASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USA
机构:
UNIV PARIS 07, INST JACQUES MONOD, INST MIXTE, CNRS, F-75251 PARIS 05, FRANCEUNIV PARIS 07, INST JACQUES MONOD, INST MIXTE, CNRS, F-75251 PARIS 05, FRANCE
BOYER, JC
HAENNI, AL
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UNIV PARIS 07, INST JACQUES MONOD, INST MIXTE, CNRS, F-75251 PARIS 05, FRANCEUNIV PARIS 07, INST JACQUES MONOD, INST MIXTE, CNRS, F-75251 PARIS 05, FRANCE
机构:
WASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USAWASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USA
CHENG, S
FOCKLER, C
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h-index: 0
机构:
WASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USAWASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USA
FOCKLER, C
BARNES, WM
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h-index: 0
机构:
WASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USAWASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USA
BARNES, WM
HIGUCHI, R
论文数: 0引用数: 0
h-index: 0
机构:
WASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USAWASHINGTON UNIV, SCH MED, DEPT BIOCHEM & MOLEC BIOPHYS, ST LOUIS, MO 63110 USA