Team:Todai-Tokyo/Notebook/bread

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=='''Plan'''==
=='''Plan'''==
'''Aim:'''Create yeast that can be used to make sweet and low energy bread<BR>  
'''Aim:'''Create yeast that can be used to make sweet and low energy bread<BR>  

Revision as of 00:47, 20 October 2009

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the notebook

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

10/1~13

  • 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

10/15

  • ligate mtlD and plate1 7D

10/17

  • colony PCR of mtlD+plate1-7D

10/18

  • Miniprep of mtlD+plate1-7D
  • MtlD made a debut as an iGEM part!





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