Difference between revisions of "Part:BBa K1049001:Experience"

Revision as of 04:17, 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. 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.

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, 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.

User Reviews

UNIQ7d4391f15d16ee70-partinfo-00000000-QINU UNIQ7d4391f15d16ee70-partinfo-00000001-QINU

@@ Line 5: / Line 5: @@
 ===Applications of BBa_K1049001===
-===Applications of BBa_K1049002===
+===Applications of BBa_K1049001===
+We constructed ATF2 generator; BBa_K1049002 in order to measure the quantitative data.
 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:KIT2013SDS.png|500px|thumb|left|Figure.1 SDS-PAGE Starting from the left, marker, ATF2 at 28 degree C, ATF2 + IPTG at 28 degree C, marker, ATF2 at 37 degree C, ATF2  + IPTG at 37 degree C]]
+[[File:KIT2013SDS.png|500px]]
 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, 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.
@@ Line 30: / Line 41: @@
 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.