Difference between revisions of "Part:BBa K3105682"

Revision as of 22:55, 19 October 2019

Expression construct MnP

Manganese Peroxidase (MnP) cloned in-frame with α-factor from Saccharomyces cerevisiae. This construct is controlled by the AOX1 promotor, which is inducible by methanol.

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 1438
Illegal BglII site found at 1730
Illegal XhoI site found at 1189
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 2491
1000
COMPATIBLE WITH RFC[1000]

Characterization

The part was expressed in X-33 P. pastoris cells using the pPICZαB shuttle vector. Cultures were induced with methanol and analysed on SDS-PAGE. MnP was successfully expressed and secreted (Figure 1).

Figure 1: Expression and secretion of Manganese Peroxidase (MnP)
X-33 P. pastoris cells were transformed with pPICZαB_MnP, induced (i), fractionated into pellets (P) and supernatants (S), and analysed by 10 % SDS-PAGE stained with Coomassie Blue. u, uninduced
Induction bands are visible at approximately 70 kDa for both P and S (red arrows), showing secretion of MnP.

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 Manganese Peroxidase (MnP) cloned in-frame with &#945;-factor from <I>Saccharomyces cerevisiae</I>. This construct is controlled by the AOX1 promotor, which is inducible by methanol.
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+===Usage and Biology===
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