"
Team:Todai-Tokyo/Notebook/bread
From 2009.igem.org
(Difference between revisions)
(→"'9/21) |
|||
| Line 4: | Line 4: | ||
'''Methods:'''<BR> | '''Methods:'''<BR> | ||
| - | #Clone the glucoamylase | + | #1.Clone the Glu1 (glucoamylase) from Saccharomycopsis fibuligera and insert it in the yeast chromosome by homologous recombination<BR> |
| - | #Clone mtlD (mannitol synthase) from E.coli and insert it in the yeast chromosome by homologous recombination | + | #2-a Clone mtlD (mannitol synthase) from E.coli and insert it in the yeast chromosome by homologous recombination |
| - | #Replace gpd1 gene by | + | #Replace gpd1 gene by mtlD and gpd2 gene by glu1<BR> |
| + | |||
| + | #2-b Clone xylose isomerase gene and D-tagatose 3-epimerase gene from Mesorhizobium loti | ||
| + | #Replace gpd1 gene by D-tagatose 3-epimerase gene and gpd2 gene by glu1. Transform a plasmid coding xylose isomerase gene. | ||
=='''~9/20'''== | =='''~9/20'''== | ||
Revision as of 03:50, 16 October 2009
| Home | The Team | The Project | Parts Submitted to the Registry | Modeling | Notebook | Protocols | Ethics |
|---|
Contents |
Plan
Aim:Create yeast that can be used to make sweet and low energy bread
Methods:
- 1.Clone the Glu1 (glucoamylase) from Saccharomycopsis fibuligera and insert it in the yeast chromosome by homologous recombination
- 2-a Clone mtlD (mannitol synthase) from E.coli and insert it in the yeast chromosome by homologous recombination
- Replace gpd1 gene by mtlD and gpd2 gene by glu1
- 2-b Clone xylose isomerase gene and D-tagatose 3-epimerase gene from Mesorhizobium loti
- Replace gpd1 gene by D-tagatose 3-epimerase gene and gpd2 gene by glu1. Transform a plasmid coding xylose isomerase gene.
~9/20
- PCR of mtlD
- PCR of gpd1 promoter
- TA cloning of mtlD
9/21
- Miniprep of mtlD
9/22
- read the sequence of mtlD→successful
- mtlD primers with HAtag come
9/25
- PCR of mtlD with new primer→failed
9/26
- PCR of mtlD with new primer→successful
9/27
- PCR of gpd1 promoter with Pfu Ultra
October
- PCR of Glu1
- TA cloning of Glu1
- PCR of gpd1 promoter with ExTaq
10/14
- cut mtlD and plate1 7D both by EcoRI and PstI
- colony PCR of Glu1
| Home | The Team | The Project | Parts Submitted to the Registry | Modeling | Notebook | Protocols | Ethics |
|---|
