Difference between revisions of "Part:BBa K1049001:Experience"
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===Applications of BBa_K1049001=== | ===Applications of BBa_K1049001=== | ||
We constructed ATF2 generator; BBa_K1049002 in order to measure the quantitative data. | We constructed ATF2 generator; BBa_K1049002 in order to measure the quantitative data. | ||
| − | + | 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:KIT2013SDS.png|500px]] | [[File:KIT2013SDS.png|500px]] | ||
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You can find the band at lanes which are added IPTG just beneath the band of 68kDa. | You can find the band at lanes which are added IPTG just beneath the band of 68kDa. | ||
| − | In addition, according to this previous study, the ability of ATF2 protein producing isoamyl acetate in yeast is higher than ATF1 protein. | + | 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. | It is known that both ATF1 and ATF2 protein are involved in producing isoamyl acetate. | ||
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Herewith, our team, KIT-Kyoto 2013 iGEM team, meets the additional requirements for a Gold Medal. | Herewith, our team, KIT-Kyoto 2013 iGEM team, meets the additional requirements for a Gold Medal. | ||
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| + | ---- | ||
| + | |||
| + | [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=== | ===User Reviews=== | ||
Revision as of 04:26, 24 September 2013
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_K1049001
We constructed ATF2 generator; BBa_K1049002 in order to measure the quantitative data. 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.
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.
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
UNIQ52f02615d79f524b-partinfo-00000000-QINU UNIQ52f02615d79f524b-partinfo-00000001-QINU