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. 2022 Nov 21;14(11):2589. doi: 10.3390/v14112589

Figure 9.

Figure 9

Recombinant phages can acquire the missing genes from the donor plasmid expressed in Pseudomonas host cells. (A) Transcripts of partial segment of L (pLM1009), isolated l segment from mutant carrying amber mutation from φ6 sus351, and complete ssRNA from m and s segments of φ6 are in vitro packaged in PC and coated with P8, transducing the spheroplasts of HB10Y with plasmid pLM1003, coding the missing 2, 4, and 7 genes and MCS of pUC8 (B). The phage φ1980 carries at least two L segments and M and S segments (C,D). On the dsRNA agarose gel picture reproduced from Onodera et al. (1995) (C), the pattern of φ1980 dsRNA shows a higher intensity of the L band compared to any of the dsRNA bands from the control φ6 dsRNA pattern. If the φ1980 was plated on HB10Y host without donor plasmid pLM1003, the extremely rare lytic plaques were all recombinants. (E) The phages packed an extra segment bearing missing 2, 4, and 7 genes and the replication signal from either the M or the S segment. The recombinants had minimal homology (the homology regions are shown as bold black symbols, whereas the recombined RNA origins are shown as light green for the L segment, light pink for S, and blue for pUC8 MCS). Based on Onodera et al. (1995).