Difference between revisions of "Part:BBa K646004"

Latest revision as of 02:29, 5 October 2011

T7 promoter with Hemeoxygenase

Using parts BBa_K646000 and BBa_I712074, a T7 promoted Hemeoxygenase was created by digesting the T7 biobrick with SpeI and PstI and ligating it to the HO insert digested with XbaI and PstI. This procedure can be used if an EcoRI and PstI cut plasmid backbone is not available.

Successful expression in E.coli should result in green colonies due to the presence of biliverdin (synthesized by heme oxygenase 1).

Usage and Biology

The BioBrick part in team Macquarie University (2011) mechanism

Liquid media containing IPTG and ALA, incubated for 24 hours at low temperature, spun down. Pellets are green due to the production of biliverdin from the breakdown of heme - this indicates that heme oxyganse was expressed using T7 promoter.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

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 [[Image:Results_-_green colonies.JPG|400px]]
-Liquid media containing IPTG and ALA, incubated for 24 hours at low temperature, spun down. Pellets are green as heme oxyganse was expressed using T7 promoter and degraded heme into biliverdin.
+Liquid media containing IPTG and ALA, incubated for 24 hours at low temperature, spun down. Pellets are green due to the production of biliverdin from the breakdown of heme - this indicates that heme oxyganse was expressed using T7 promoter.
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