4 mM IPTG
with post induction incubation period of 6h at 37AdegC.
coli Rosseta, we optimized expression temperature, IPTG
induction concentration and time.
coli Expression Systems including, Sugar Inducible, Depletion Inducible, Optimized IPTG
Inducible Platforms, and Lonza s pDNA Services Platform.
coli DH5 transgenicas mediante induccion con IPTG
, demostrado por los valores obtenidos con la tecnica de BCA de 1.
0, and induced with 60 [micro]M IPTG
(isopropyl-1-thio-,3-D-galactoside) for 3 h at 37[degrees]C.
Due to IPTG
induction, white colored colonies containing cellulase gene were produced after overnight incubation at 37 @C.
After induction with 1 mM IPTG
for 2 h at 37[degrees]C, bacteria BL21 (DE3) transformed with pET28a (+)-endog expresses a protein of approximately 45 kDa, corresponding to the expected size of the rENDOG, was observed (Figure 2A).
Neste ponto, a expressao da CP foi induzida por adicao de IPTG
(2mM) e, 6h apos a inducao, as celulas foram coletadas por centrifugacao (6000g por 10min).
The proteins (NT-gp96 and CT-gp96) were induced individually using 1 mM IPTG
This gene can be actively transcribed to produce [beta]-galactosidase with the presence of an activator, IPTG
Autoclave to sterilize; cool to around 50[degrees]C; add appropriate antibiotics and other supplements, such as X-Gal and IPTG
, if needed; and then pour).
The ligated cDNA were precipitated, cloned into UniZAP XR (Stratagene) vector at EcoR I site and plated on XL1 Blue cells in the presence of X-gal and IPTG