Expression of a Peptidoglycan Hydrolase from Lytic Bacteriophages Atu_ph02 and Atu_ph03 Triggers Lysis of Agrobacterium tumefaciens

Supplemental material

  • Supplemental file 1 -

    Supplemental methods: clonal isolation of bacteriophage strains, concentration and partial purification of virions, preparation of virion DNA, DNA restriction analysis; comparison of gene products encoded in Atu_ph02 and Atu_ph03 (Table S1); similarity of putative Atu_ph03 proteins to proteins in select bacteriophages (Table S2); initial characterization of phage genomic DNA shows that Atu_ph02 and Atu_ph03 are distinct (Fig. S1); phages Atu_ph03 and Atu_ph02 are very similar (Fig. S2); Clustal alignment of DUF3380 domains from various phage proteins with similarity to the DUF3380 from Salmonella phage 10 endolysin and bacterial PG-binding proteins with similarity to the DUF3380 found in PPH (Fig. S3); growth of A. tumefaciens with plasmids to express variants of pph under uninduced conditions (Fig. S4); clearing of peptidoglycan is observed when PPH is expressed in A. tumefaciens (Fig. S5); legends to supplemental movies.

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  • Supplemental file 2 -

    Time-lapse microscopy in DIC of A. tumefaciens strain C58 growing for 7 h (Movie S1).

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  • Supplemental file 3 -

    Time-lapse microscopy in DIC of cells infected with Atu_ph03 at an MOI of 0.01 for 7 h (Movie S2).

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    Time-lapse microscopy in DIC of cells infected with Atu_ph02 at an MOI of 0.01 for 7 h (Movie S3).

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    Time-lapse microscopy in DIC of cells expressing pSRKKm-Plac-sfgfp for 16 h (Movie S4).

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    Time-lapse microscopy in DIC of cells expressing pSRKKm-Plac-PPH growing for 16 h (Movie S5).

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    Time-lapse microscopy in DIC of cells expressing pSRKKm-Plac-PPHE32A growing for 20 h (Movie S6).

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    Time-lapse microscopy in DIC of cells expressing pSRKKm-Plac-PPHK328A,K334A,K335A growing for 7 h (Movie S7).

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    Time-lapse microscopy in DIC of cells expressing pSRKKm-Plac-PPHE32A,K328A,K334A,K335A growing for 16 h (Movie S8).

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  • Supplemental file 10 -

    Time-lapse microscopy in DIC of cells expressing pSRKKm-Plac-PPH1-304 growing for 16 h (Movie S9).

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