Coding

Part:BBa_K4278702

Designed by: Shi Lifang   Group: iGEM22_Shanghai_Metropolis   (2022-09-20)


ATF1

ATF1

Profile

Name: ATF1

Base Pairs: 1575 bp

Origin: Saccharomyces cerevisiae, genome

Properties: the alcohol acetyltransferase

Usage and Biology

This part is a basic part named ATF1, and the ATF1 gene encodes a single alcohol acetyl-CoA transferase (AATase) enzyme, because of the importance of the ATF1 in the aroma profile of the wine, its regulation during fermentation has been investigated in detail. The yeast with a constitutively overexpressed ATF1 gene produces more isoamyl acetate and ethyl acetate compared to a wild-type strain [1-5]. The ATF1 enzyme contains transmembrane domains which fix the protein in the membrane of lipid droplets that bud out from the endoplasmic reticulum (ER). This enzyme widely exists in different varieties of yeast.

Construct design

The ATF1 gene is amplified from the Saccharomyces cerevisiae genome by PCR and was used for further research.

Experimental approach

1. ATF1 Gel Electrophoresis

Figure 1. Figure 1. The PCR gel electrophoresis result of the Kana gene fragment. M. DNA marker, 1. The ENA fragment of ATF1.

In order to obtain our target genes, we amplified the target gene ATF1 from the Saccharomyces cerevisiae genome. We did gel electrophoresis to verify if we successfully amplified the target fragment. In figure1, a clear and single DNA band could be seen, indicating that we successfully amplified our target genes. We extracted the DNA fragments and stored them at -20℃ for future use.

References

1.Zhang J., Zhang C., Wang J., Dai L., Xiao D. (2014) Expression of the Gene Lg-ATF1 Encoding Alcohol Acetyltransferases from Brewery Lager Yeast in Chinese Rice Wine Yeast. In: Zhang TC., Ouyang P., Kaplan S., Skarnes B. (eds) Proceedings of the 2012 International Conference on Applied Biotechnology (ICAB 2012). Lecture Notes in Electrical Engineering, vol 249. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-37916-1_5.

2.Ma L, Huang S, Du L, Tang P, Xiao D. Reduced Production of Higher Alcohols by Saccharomyces cerevisiae in Red Wine Fermentation by Simultaneously Overexpressing BAT1 and Deleting BAT2. J Agric Food Chem. 2017 Aug 16;65(32):6936-6942. doi: 10.1021/acs.jafc.7b01974.

3.Lilly M, Bauer FF, Styger G, Lambrechts MG, Pretorius IS. The effect of increased branched-chain amino acid transaminase activity in yeast on the production of higher alcohols and on the flavour profiles of wine and distillates. FEMS Yeast Res. 2006 Aug;6(5):726-43. doi: 10.1111/j.1567-1364.2006.00057.x.

4.Dai L., Zhang C., Zhang J., Qi Y., Xiao D. (2014) Effects of IAH1 Gene Deletion on the Profiles of Chinese Yellow Rice Wine. In: Zhang TC., Ouyang P., Kaplan S., Skarnes B. (eds) Proceedings of the 2012 International Conference on Applied Biotechnology (ICAB 2012). Lecture Notes in Electrical Engineering, vol 249. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-37916-1_42.

5.Choi YJ, Lee J, Jang YS, Lee SY. Metabolic engineering of microorganisms for the production of higher alcohols. mBio. 2014 Sep 2;5(5):e01524-14. doi: 10.1128/mBio.01524-14.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 96
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 523


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