Generator

Part:BBa_K1049002:Experience

Designed by: Yohei TAMIAKI   Group: iGEM13_KIT-Kyoto   (2013-09-19)
Revision as of 04:29, 24 September 2013 by Okaeri007 (Talk | contribs) (→‎Applications of BBa_K1049002)


This experience page is provided so that any user may enter their experience using this part.
Please enter how you used this part and how it worked out.

Applications of BBa_K1049002

Our team KIT-Kyoto 2013 constructed this isoamyl acetate generator 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.

KIT2013SDS.png

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.

In addition, according to this previous study [1], the ability of ATF2 protein producing isoamyl acetate in yeast is higher than ATF1 protein.

It is known that both ATF1 and ATF2 protein are involved in producing isoamyl acetate.

Earlystudy.png

In 2006, MIT iGEM team submitted ATF1 coding sequence. (BBa_J45006)

Our new part, ATF2 coding sequence, fall under the category of the improvement of function existing BioBrick Part, BBa_J45006.

Herewith, our team, KIT-Kyoto 2013 iGEM team, meets the additional requirements for a Gold Medal.

[1] Yoshimoto Hiroyuki et al. "Mechanisms of Acetate Ester Production and Control in Yeasts -Monograph-", seibutsu-kogaku kaishi 79(2), 33-40, 2001-02-25

User Reviews

UNIQf6b5a3f16ca6ad64-partinfo-00000000-QINU UNIQf6b5a3f16ca6ad64-partinfo-00000001-QINU