Difference between revisions of "Part:BBa K1403019:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | This sequence was submitted downstream a standard T7 constitutive promoter inside a | + | This sequence was submitted downstream a standard T7 constitutive promoter inside a pETDUET vector. |
The ilvBN sequence was isolated from MG1655 through PCR and digested with NCoI and EcoRI to put into pETDUET1 vector. It was transformed into E.Coli strain with plasmid containing aldB to produce acetoin, which has the smell of butter. | The ilvBN sequence was isolated from MG1655 through PCR and digested with NCoI and EcoRI to put into pETDUET1 vector. It was transformed into E.Coli strain with plasmid containing aldB to produce acetoin, which has the smell of butter. | ||
| − | + | We chose pETDUET1, instead of PSB1C3, because this vector has promoter in itself and makes the cloning process easier. This part is not necessary for production of any acetoin, but important for a high amount. | |
===Source=== | ===Source=== | ||
Revision as of 14:58, 12 October 2014
Acetohydroxybutanoate synthase /acetolactate synthase (ilvB/N) expression cassette
- 10INCOMPATIBLE WITH RFC[10]Illegal XbaI site found at 47
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal XbaI site found at 47
- 25INCOMPATIBLE WITH RFC[25]Illegal XbaI site found at 47
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This sequence was submitted downstream a standard T7 constitutive promoter inside a pETDUET vector.
The ilvBN sequence was isolated from MG1655 through PCR and digested with NCoI and EcoRI to put into pETDUET1 vector. It was transformed into E.Coli strain with plasmid containing aldB to produce acetoin, which has the smell of butter.
We chose pETDUET1, instead of PSB1C3, because this vector has promoter in itself and makes the cloning process easier. This part is not necessary for production of any acetoin, but important for a high amount.
Source
E. coli MG1655
References
David R. Nielsen, Sang-Hwal Yoon, Clara J. Yuan and Kristala L. J. Prather, Metabolic engineering of acetoin and meso-2,3-butanediol biosynthesis in E. coli, January 2010.