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- W1567281123 abstract "This chapter focuses on the reverse genetics of double-stranded RNA (dsRNA) bacteriophage Φ6. Φ6 is a bacteriophage that infects the plant pathogen Pseudomonas syringae pv phaseolicola. It has a unique structure and life cycle in that it is the only bacteriophage with a genome of dsRNA and the only phage that has a lipid-containing membrane that surrounds a polyhedral nucleocapsid. The virion contains three segments of dsRNA. The chapter discusses life cycle of Φ6 infection. The second method of reverse genetics is done in vivo. A culture of P. syringae––that is, carrying a plasmid that produces a transcript with imprecise ends in which is embedded a complete copy of one of the genomic segments––is prepared. These cells are infected with a derivative of Φ6 that contains a deletion in the segment to be exchanged. In vitro genomic packaging with bacteriophage and reverse genetics by in vivo transcript acquisition is also presented in this chapter. Using these two methods, novel genes can be inserted into the 3´ noncoding regions of the three segments. Genes are placed for kanamycin resistance (aph), for the α portion of β-galactosidase (lacH), and for green fluorescent protein GFP (gfp). Nonsense mutations in genes are engineered (e.g., genes 10 and 14) to assess their function. Virus-containing genomic segments with small or large deletions or virus with segments joined together so that the number of genomic segments can be two or even one are prepared." @default.
- W1567281123 created "2016-06-24" @default.
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- W1567281123 date "1999-01-01" @default.
- W1567281123 modified "2023-09-25" @default.
- W1567281123 title "Reverse Genetics of dsRNA Bacteriophage Φ6" @default.
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- W1567281123 doi "https://doi.org/10.1016/s0065-3527(08)60355-3" @default.
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