Extended Data Fig. 3: AvcD C-terminal 6x Histidine fusion maintains the same activity as the WT AvcD enzyme and the presence of avcI does not reduce the abundance of AvcD.
From: Phage defence by deaminase-mediated depletion of deoxynucleotides in bacteria
(A) Representative images of WT V. cholerae and Δig222 cultures maintaining an empty vector plasmid (pVector1) or Ptac-inducible avcD-6xHIS plasmid (pAvcD6xHis) grown in the presence of 100 µM IPTG for 2 h. Cells were stained with FM4-64 prior to imaging and performed in biological triplicate. (B) Representative coomassie stained PAGE gel (top) and matched anti-6x His antibody Western blot (bottom) of whole cell lysates normalized to total protein from V. cholerae WT and Δig222 cultures maintaining pVector1 or pAvcD6xHis. Black triangles correspond to AvcD6xHis (60.6 kDa). Analysis was performed in biological triplicate and the relative signal intensity (C) was the determined by comparing the intensities of AvcD6xHis from paired WT and Δig222 lysates probed on the same blots. Data represent the mean ± SEM of three biological replicate. Statistical significance was determined using two-sided Student’s t-test. P values between WT and Δig222 is 0.185. ns indicate not significant.
