Team:UT-Dallas/Project/results

RESULTS

a) Fluorescence microscopy of Escherchia coli cells. Cells were transformed with a plasmid that produces the Streptococcus pyogenes Cas9 protein under the control of the pTet promoter and a constitutive gRNA that targets the Vibrio cholerae gene acfA and a plasmid that produces a yellow fluoescence reporter with a 20 base pair sequence from the Vibrio cholerae gene acfA. The first column is a bright field image, the second column is the yellow fluorescence channel, and the third column is the yellow channel with color. b) Fluorescence microscopy of Escherchia coli cells. Cells were transformed with a plasmid that produces the Streptococcus pyogenes Cas9 protein under the control of the pTet promoter and a plasmid that produces a yellow fluoescence reporter with a 20 base pair sequence from the Vibrio cholerae gene acfA. This is a control for panel (a), having no gRNA. The first column is a bright field image, the second column is the yellow fluorescence channel, and the third column is the yellow channel with color. C) Serial dilution of the cells in panels (a) & (B). The cells were dilued down to 1/10,000,000 and plated on carbenicillin and chlorampenicol plates. d) Graph showing colonies versus dilution factor from the serial dilution in panel (C).