Table 2.

Culture absorbance change after induction of plasmid-expressed D-fusion genes. ^a

Plasmid In E. coli, Encoded D or D-Fusion Protein	Non-Induces (0 Time)	Non-Induced, 140 min Cell Growth at 30 °C	Induced, 140 min Cell Growth after Shifting Culture to 42 °C
p613, gpDcoe	0.10	0.44	0.79
p627, gpD-PR39	0.14	0.51	0.70
p625, gpD-LL37	0.15	0.71	0.70

^a Single colony isolates of strains p613, p625 and p627 (refer to Table 1 and Table 3 and Section 2.3), respectively 594[pcIpR-Dcoe-timm], 594[pcIpR-Dcoe-PR39-timm] and 594[pcIpR-Dcoe-LL37-timm], were inoculated into LB-Amp50 medium and grown up in a 30 °C shaking water bath until reaching an A₅₇₅ of about 0.1. Each culture was split: One-half was returned to the 30 °C shaking bath to maintain repression of the plasmid cloned GOI. The parallel culture was shaken for 15 s at 60 °C (this rapidly brings the culture temperature up to ~39 °C) and then shaken in a 42 °C bath to permit full expression of the plasmid encoded GOI. The A₅₇₅ values were recorded at 60, 90, 120 and 140 min (see Figure S2).