Difference between revisions of "Part:BBa K1403019:Design"

Latest revision as of 07:41, 31 October 2014

Acetohydroxybutanoate synthase /acetolactate synthase (ilvB/N) expression cassette

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal XbaI site found at 47
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
INCOMPATIBLE WITH RFC[23]
Illegal XbaI site found at 47
25
INCOMPATIBLE WITH RFC[25]
Illegal XbaI site found at 47
1000
COMPATIBLE WITH RFC[1000]

Design Notes

The purpose of this part is to overexpress ilvB/N in E. coli.

The ilvB/N genes were isolated from E. coli MG1655 through PCR, digested with NcoI and EcoRI, and cloned into the pETDUET-1 vector downstream a T7 promoter and RBS. This part is not strictly necessary for production of acetoin in E. coli but improves the production thereof.

Oligos:

Fwd: 5'- GTTCAAGCCTTGAGCGGTTACTG -3'

Rev: 5'- GAACTTGCCATGCTCCAGTCCT -3'

Source

ilvB/N coding sequences from E. coli MG1655 ([http://www.ncbi.nlm.nih.gov/nuccore/545778205?from=3850802&to=3851092&sat=4&sat_key=117098803&report=gbwithparts U00096.3])

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.

@@ Line 8: / Line 8: @@
 The purpose of this part is to overexpress <i>ilvB/N</i> in <i>E. coli</i>.
-The <i>ilvB/N</i> genes were isolated from <i>E. coli</i> MG1655 through PCR, digested with NcoI and EcoRI and cloned into the pETDUET-1 vector downstream a T7 promoter and RBS.
+The <i>ilvB/N</i> genes were isolated from <i>E. coli</i> MG1655 through PCR, digested with NcoI and EcoRI, and cloned into the pETDUET-1 vector downstream a T7 promoter and RBS.
-This part is not strictly necessary for production of acetoin in <i>E. coli</i>, but it improves its production.
+This part is not strictly necessary for production of acetoin in <i>E. coli</i> but improves the production thereof.
 Oligos: