Additional file 1 of Variants of a putative baseplate wedge protein extend the host range of Pseudomonas phage K8

Additional file 1: Supplementary Table S1. Bacteria, plasmids, and phages used in this study. Supplementary Table S2. Primers used in this study. Supplementary Table S3. Molar mass of GP075 and its derivatives. Supplementary Table S4. Possible protein interactions identified by the bacterial two-hybrid system a. Supplementary Table S5. Mutations of the gp075 gene of the 41 isolated K8 mutants a. Supplementary Figure S1. Protein structure modelling by using the online server SWISS-MODEL. The amino acid sequences of the wild-type protein GP075 and its two variants GP075m and GP075-D7 were uploaded for structural template searching and model building. The upper panels showed the brief descriptions of the secondary structure parameters of each protein. The bottom panels displayed the 3D structure images of the indicated proteins. Supplementary Figure S2. SDS-PAGE of the purified viral proteins. GP076: 670 amino acids. GP075: 243 amino acids. GP075m: GP075 with one amino acid substitution T239A. GP075-D7: GP075 with one copy of the 7-aa duplication (APWYSVG). GP075-D14: GP075 with two identical copies of the 7-aa duplication. GP075-D21: GP075 with three identical copies of the 7-aa duplication. M: Premixed Protein Marker (Low) from Takara. Supplementary Figure S3. Native-PAGE of the purified viral proteins. GP075: 243 amino acids. GP075m: GP075 with one amino acid substitution T239A. The protein samples for electrophoresis were treated before loading as follows: 1 and 5, the protein samples were mixed with 2×loading buffer (without SDS); 2 and 6, the protein samples were mixed with 2×loading buffer (without SDS) and incubated at 100°C for 10 min. 3 and 7: the protein samples were mixed with 2×loading buffer (containing 4% SDS). 4 and 8: the protein samples were mixed with 2×loading buffer (containing 4% SDS) and incubated at 100°C for 10 min. M: Premixed Protein Marker (Low) from Takara. Supplementary Figure S4. Thermostability of the K8 mutants. a. Amino acids sequences alignments of the GP075-like proteins from the isolated K8 mutants. Red letters represent the amino acids duplication or the substitutions. Numbers represent the positions of amino acid residuals in the GP075 protein. b. Thermostability of the K8 mutants. Phages were treated at various temperatures for 1 h before determination of the live phages. The phage titers obtained at the treatment at 4°C were used as controls to calculate the survival percentages. The experiments were independently replicated three times. One-way ANOVA followed by a Tamhane T2 test was performed to compare the means of the groups treated at 70°C. Supplementary Dataset 1. LC-MS data. Supplementary Dataset 2. Ampliconsequencing data. Supplementary Dataset 3.Homologs of GP075.