Table 2.

Expression of a C-terminally Ano-EGFP fusion protein.

Sample	Fluorescence 0 mM IPTG	Fluorescence 10 mM IPTG	Significance
E. coli Top10 + pEGFP-ano-Start1	0	0	N/A
E. coli Top10 + pEGFP-ano∗-Start1	0	1124 ± 152	N/A
E. coli Top10 + pEGFP-ano-Start2	0	0	N/A
E. coli Top10 + pEGFP-ano∗-Start2	0	0	N/A
E. coli Top10 + pEGFP-ano-Start3	0	0	N/A
E. coli Top10 + pEGFP-ano∗-Start3	0	291 ± 56	N/A
E. coli Top10 + pEGFP-ano-Start5	883 ± 703	3305 ± 236	∗∗∗
E. coli Top10 + pEGFP-ano∗-Start5	466 ± 312	397 ± 169	–
E. coli Top10 + pEGFP-ECs2385	527 ± 430	8662 ± 2444	∗∗∗

Escherichia coli Top10 was transformed with the different pEGFP-ano plasmids and expression of the fusion protein was induced using 10 mM IPTG. OD₆₀₀ was adjusted to 0.6 and fluorescence was measured. Fluorescence values of empty E. coli Top10 were subtracted (causing zero values for some readings). The experiment was performed in triplicate. The first three potential start codons had zero fluorescence, when not induced. Therefore, calculation of significance was meaningless (N/A) for those. Significant changes between induced and uninduced cultures were calculated by two-tailed Student’s t-test (^∗∗∗p < 0.001).