Difference between revisions of "Part:BBa K3239007"

Revision as of 10:54, 15 October 2019

pAOX1-GFP

yEFGP3 reporter gene expressed by pAOX1

Usage and Biology

This construct is the control used in our experiment. After homologous recombination into wild type P. pastoris, the EGFP expressed shall serve as an indicator of AOX1 expression levels. Note that the original pAOX1-AOX1 copy remains intact after the recombination.

Experience

we labeled our strains containing this construct AOX1-GFP and used it as the positive control of our data. The following data are acquired after 108h YNM (1.34% yeat nitrogenous base with 0.5% methanol, 50mg/mL histidine and 10mg/mL biotin) incubation, sampled every 12h. Methanol is added to the media every 24 hours to maintain the methanol concentration. Three parallels are performed and the data shown is the average of the three parallels.

Figure 1. AOX1-GFP methanol consumption curve

Source

Both the promoter and the reporter are registered. pAOX1: BBa_K3239003 yEGFP3: BBa_K3239006

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 935
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 5: / Line 5: @@
 yEFGP3 reporter gene expressed by pAOX1
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
-This is one of our constructs aimed at moderately upregulating pAOX1 activity in P.pastoris GS115. After homologous recombination into wild type P. pastoris, the EGFP expressed shall serve as an indicator of AOX1 expression levels. Note that the original pAOX1-AOX1 copy remains intact after the recombination.
+This construct is the control used in our experiment. After homologous recombination into wild type P. pastoris, the EGFP expressed shall serve as an indicator of AOX1 expression levels. Note that the original pAOX1-AOX1 copy remains intact after the recombination.
+===Experience===
+we labeled our strains containing this construct AOX1-GFP and used it as the positive control of our data.
+The following data are acquired after 108h YNM (1.34% yeat nitrogenous base with 0.5% methanol, 50mg/mL histidine and 10mg/mL biotin) incubation, sampled every 12h.
+Methanol is added to the media every 24 hours to maintain the methanol concentration.
+Three parallels are performed and the data shown is the average of the three parallels.
+[[File:Methanol(24h)AOX1-GFP.png|800px|thumb|left|Figure 1. AOX1-GFP methanol consumption curve]]
+===Source===
 Both the promoter and the reporter are registered.
 pAOX1: BBa_K3239003