Team:Todai-Tokyo/Notebook/bioclock

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= Plan =
= Plan =
'''Aim:''' Create E.coli that show oscillatory gene expression pattern
'''Aim:''' Create E.coli that show oscillatory gene expression pattern

Revision as of 00:47, 20 October 2009

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the notebook

Contents

Plan

Aim: Create E.coli that show oscillatory gene expression pattern (colE1 ori vector)-placI/araC-araC+ssra tag-double terminator-placI/araC-GFP+ssra tag-double terminator- (p15A ori)-placI/araC-lacI+ssra tag-double terminator-

Methods:
1. Clone the following genes from Bacillus subtilis into a biobrick vector.


  1. plate1,13B
  2. plate1,13L
  3. plate2,1H

2. Make a gene network that express oscillatory pattern, using these genes.


7/7

Cloning the parts
preculture of the Biobrick parts for Miniprep

7/8

Cloning the parts
Miniprep of E.coli cells containing Biobrick parts with Promega, Wizard Plus SV Miniprep DNA Purification System

failure

7/9

Cloning the parts
Miniprep again of E.coli cells containing Biobrick parts with Promega, Wizard Plus SV Miniprep DNA Purification System

7/29

Miniprep

  1. P1.14L(araC)
  2. P1.7L(lacI)
  3. P1.4E(cI)
  4. P1.3D(ColE1)
  5. P1.9C(p15A)
  6. P1.9G(p15A)

7/30

Miniprep

  1. P1.14L(araC)
  2. P1.7L(lacI)
  3. P3.21D

August/September

  • create plasmid including EcoRI restriction site, XbaI restriction site, placI/araC, XhoI restriction site, NcoI restriction site, double terminator, SpeI restriction site and PstI restriction site.→pUC57

-E-X-placI/araC-XhoI-NcoI-double terminator-S-P-

  • cut pUC57 by XhoI and NcoI

→insert GFP, araC or lacI
(GFP:2006 plate1-16E,araC and lacI:2009 iGEM distribution)

  • cut plate1-23L(double terminator) and insert plate1-7L(lacI) in it
  • PCR of cI~cII

October

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