View source for Part:BBa K1049002:Experinece

Our team KIT-Kyoto 2013 constructed this part for the purpose of measurement. T7 promoter is an IPTG-inducible promoter. We added 20uL IPTG (100mM) to our genetically modified E.coli after cultivation at 28 and 37 degree C. 2 hours after, we extracted soluble proteins from it by using FastBreak™ Cell Lysis Reagent and did SDS-polyacrylamide gel electrophoresis.
[[File:ATF2SDS.png|500px|thumb|left|Figure.1 SDS-PAGE Starting from the left, marker, candidate 1 at 28 degree C, candidate 1 + IPTG at 28 degree C, candidate 2 at 28 degree C, candidate 2 + IPTG at 28 degree C, marker, candidate 1 at 37 degree C, candidate 1 + IPTG at 37 degree C, candidate 2 at 37 degree C, candidate 2 + IPTG at 37 degree C.
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ATF2 gene encodes AATase, which is about 62kDa. The consumption of protein marker is like this.

Myosin                                 200kDa
β‐Galactosidase                   120kDa
Bovine Serum Albumin         95kDa
Glutamine dehydrogenase    68kDa
Ovalbumin                           50kDa
Carbonic Anhydrase             36kDa
Myoglobin                            27kDa
Lysozyme                             20kDa
Aprotinin                              10kDa
 
You can find the band at lanes which are added IPTG just beneath the band of 68kDa.