Rickettsia typhi Possesses Phospholipase A2 Enzymes that Are Involved in Infection of Host Cells
Figure 5
Cytotoxicity in yeast strain INVSc1 by CFU assay.
The yeast strain INVSc1 transformed with the plasmids carrying genome sequence of pat1 (Pat1-wt), carrying codon optimized sequence of pat1 (Pat1-co) or carrying mutation at catalytic sites of pat1-co (Pat1-SD). A) The percentage of CFU in transformed yeast cells was determined as described (see Materials and Methods). B) Western blot analysis of the expression of Pat1 and its mutant derivatives in INVSc1 under inducing condition (SC-U+Gal medium). The total proteins from yeast cells carrying the appropriate plasmid were probed with anti-Pat1 antibody using WesternBreeze chemiluminescent immunodetection kit (Invitrogen). LaneM: MagicMark XP Western Protein Standard (Invitrogen), Lane1: Pat1-SD/INVSc1, Lane2: Pat1-co/INVSc1, Lane3: Pat1-wt/INVSc1. The size of the expected recombinant protein (including C-terminal V5 epitope and 6× His tag) is mentioned on the left (59.46 kDa). The bands below the expected protein size may have resulted from degradation of recombinant protein or nonspecific binding to proteins from yeast cell lysate.