Fig. 3: Complementation of a T1-PKS UV-mutant and results from gradient ultracentrifugation of the LINA particle. | The ISME Journal

Fig. 3: Complementation of a T1-PKS UV-mutant and results from gradient ultracentrifugation of the LINA particle.

From: Ice nucleation in a Gram-positive bacterium isolated from precipitation depends on a polyketide synthase and non-ribosomal peptide synthetase

Fig. 3

A Freezing spectra based on three replicates of droplet freezing assays (using 30 drops per dilution per replicate) of Lp VT1065 wild-type (Lp wt), Lp mutant UV14 with two SNPs in the T1-PKS gene containing the empty vector pHY300PLK, and Lp mutant UV14 complemented with plasmid pHY300PLK containing the T1-PKS gene (pT1-PKS). The X-axis shows the temperature and Y-axis shows the concentration of ice nucleation particles (INP) per colony forming unit (CFU). B The LINA particle was separated from Lp VT1065 cells and concentrated by differential filtration in a iodixanol gradient and then subjected to gradient ultracentrifugation. Samples from the indicated fractions were diluted 1:100 (to reduce the concentration of iodixanol) and analyzed by a droplet freezing assay at −11 °C (at which the difference in the number of frozen droplets was the highest). The whole experiment was repeated twice. The number of frozen droplets out of a total of 8 droplets per fraction is listed.

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