Parts Overview
As we mentioned in the description and design section, we need to build a system that can degrade pectin and lignin. And for that, we found 17 basic parts, and constructed 19 composite parts. These parts are transformed into expression vector, expressed enzyems and achieved our goals .
Parts Numbers
Parts Numbers
Parts Numbers
Basic parts
This table shows some of our basic parts, which will be used to form composite parts and build a system that can achieve our goals.
Name
Type
Coding
Coding
Coding
Coding
Coding
Coding
Coding
Coding
Coding
Coding
Description
SLAC
pelA
VP
FLO10
FLO10 pro
PHO1
PHO1 pro
SUC2
Alkali
α-factor
disigner
Hao Qiu
Hao Qiu
Hao Qiu
Hao Qiu
Hao Qiu
Hao Qiu
Hao Qiu
Hao Qiu
Hao Qiu
Hao Qiu
Length
960
1110
1101
95
294
75
276
69
384
278
Composite parts
Parts Numbers
Name
AOX1-Kozak-FLO10-SLAC-His tag-AOX1 Terminator
AOX1-Kozak-FLO10 pro-SLAC-His tag-AOX1 Terminator
AOX1-Kozak-PHO1-SLAC-His tag-AOX1 Terminator
AOX1-Kozak-PHO1 pro-SLAC-His tag-AOX1 Terminator
AOX1-Kozak-SUC2-SLAC-His tag-AOX1 Terminator
AOX1-Kozak-FLO10-pelA-His tag-AOX1 Terminator
AOX1-Kozak-FLO10 pro-pelA-His tag-AOX1 Terminator
AOX1-Kozak-PHO1-pelA-His tag-AOX1 Terminator
AOX1-Kozak-PHO1 pro-pelA-His tag-AOX1 Terminator
AOX1-Kozak-SUC2-pelA-His tag-AOX1 Terminator
AOX1-Kozak-FLO10-VP-His tag-AOX1 Terminator
AOX1-Kozak-FLO10 pro-VP-His tag-AOX1 Terminator
AOX1-Kozak-PHO1-VP-His tag-AOX1 Terminator
AOX1-Kozak-PHO1 pro-VP-His tag-AOX1 Terminator
AOX1-Kozak-SUC2-VP-His tag-AOX1 Terminator
Description
AOX1 promoter starts gene expression, signal peptide FLO10 facilitates the transport of enzymes across membranes, and His tag is used to purify SLAC enzyme.
AOX1 promoter starts gene expression, signal peptide FLO10 pro facilitates the transport of enzymes across membranes, and His tag is used to purify SLAC enzyme.
AOX1 promoter starts gene expression, signal peptide PHO1 facilitates the transport of enzymes across membranes, and His tag is used to purify SLAC enzyme.
AOX1 promoter starts gene expression, signal peptide PHO1 pro facilitates the transport of enzymes across membranes, and His tag is used to purify SLAC enzyme.
AOX1 promoter starts gene expression, signal peptide SUC2 facilitates the transport of enzymes across membranes, and His tag is used to purify SLAC enzyme.
AOX1 promoter starts gene expression, signal peptide FLO10 facilitates the transport of enzymes across membranes, and His tag is used to purify pelA enzyme.
AOX1 promoter starts gene expression, signal peptide FLO10 pro facilitates the transport of enzymes across membranes, and His tag is used to purify pelA enzyme.
AOX1 promoter starts gene expression, signal peptide PHO1 facilitates the transport of enzymes across membranes, and His tag is used to purify pelA enzyme.
AOX1 promoter starts gene expression, signal peptide PHO1 pro facilitates the transport of enzymes across membranes, and His tag is used to purify pelA enzyme.
AOX1 promoter starts gene expression, signal peptide SUC2 facilitates the transport of enzymes across membranes, and His tag is used to purify pelA enzyme.
AOX1 promoter starts gene expression, signal peptide FLO10 facilitates the transport of enzymes across membranes, and His tag is used to purify VP enzyme.
AOX1 promoter starts gene expression, signal peptide FLO10 pro facilitates the transport of enzymes across membranes, and His tag is used to purify VP enzyme.
AOX1 promoter starts gene expression, signal peptide PHO1 facilitates the transport of enzymes across membranes, and His tag is used to purify VP enzyme.
AOX1 promoter starts gene expression, signal peptide PHO1 pro facilitates the transport of enzymes across membranes, and His tag is used to purify VP enzyme.
AOX1 promoter starts gene expression, signal peptide SUC2 facilitates the transport of enzymes across membranes, and His tag is used to purify VP enzyme.
We constructed these pathways in experiments, which on the one hand enabled us to measure the effects of enzymes, and on the other hand facilitated us to select specific combinations of signal peptides and enzymes to build our system.
Name
pGAP-Kozak-FLO10 pro-pelA-His tag-TT-Pgpda-Kozak-FLO10-SLAC-His tag-TT-Pgas-Alkali-TT
pGAP-Kozak-PHO1-pelA-His tag-TT-Pgpda-Kozak-FLO10-SLAC-His tag-TT-Pgas-Alkali-TT
Pgpda-GFP
Pgas-GFP
Description
The pH regulated expression system.
The pH regulated expression system.
We used green fluorescent protein to detect the working effect of Pgpda promoter.
We used green fluorescent protein to detect the working effect of Pgas promoter.
Parts collection
Parts Numbers
Name
AOX1-Kozak-FLO10-SLAC-His tag-AOX1 Terminator
AOX1-Kozak-FLO10 pro-SLAC-His tag-AOX1 Terminator
AOX1-Kozak-PHO1-SLAC-His tag-AOX1 Terminator
AOX1-Kozak-PHO1 pro-SLAC-His tag-AOX1 Terminator
AOX1-Kozak-SUC2-SLAC-His tag-AOX1 Terminator
AOX1-Kozak-FLO10-pelA-His tag-AOX1 Terminator
AOX1-Kozak-FLO10 pro-pelA-His tag-AOX1 Terminator
AOX1-Kozak-PHO1-pelA-His tag-AOX1 Terminator
AOX1-Kozak-PHO1 pro-pelA-His tag-AOX1 Terminator
AOX1-Kozak-SUC2-pelA-His tag-AOX1 Terminator
AOX1-Kozak-FLO10-VP-His tag-AOX1 Terminator
AOX1-Kozak-FLO10 pro-VP-His tag-AOX1 Terminator
AOX1-Kozak-PHO1-VP-His tag-AOX1 Terminator
AOX1-Kozak-PHO1 pro-VP-His tag-AOX1 Terminator
AOX1-Kozak-SUC2-VP-His tag-AOX1 Terminator
pGAP-Kozak-FLO10 pro-pelA-His tag-TT-Pgpda-Kozak-FLO10-SLAC-His tag-TT-Pgas-Alkali-TT
pGAP-Kozak-PHO1-pelA-His tag-TT-Pgpda-Kozak-FLO10-SLAC-His tag-TT-Pgas-Alkali-TT
Pgpda-GFP
Pgas-GFP
By fusing enzymes with different peptides, this part collection is able to select the most suitable signal peptide for the secretion of certain types of genes, achieving the maximum efficiency and balanced protein secretion. PH sensitive promoter combined with the most efficient secretion part, the pH regulated expression system continues the former one, which can express certain enzyme as much as possible at the pH we expect.
