"

From 2011.igem.org

Danyel

• Overview
• To do list

Overview

The constructs involved in the tRNA project so far:

• General information : all values are APPROX.
  • tRNA = 100 bp
  • T7 amber = 2.7 kb
  • GFP amber = 720 bp
  • pHyperspank = 101 bp
  • Pveg_spoVG = 120 bp
  • pT7_RBS_GFP_TT = 823 bp

• tRNA amber suppressor
  • tRNA

     S38: in pSB1C3

• tRNA_TT

     S58: in pSB1C3

• Pveg_spoVG_tRNA

     S38 in S24 --- [in progress]

• Pveg_spoVG_tRNA_TT

     S81 :  this contruct in pSB1AK3 (ampR)
     S92 :  this contruct in pHM3 (ampR/tetR) --- sequencing required and transformation in subtilis attempted by Adrien.

• pHyperspank_tRNA

     S38 in S82 --- [in progress]

• pHyperspank_tRNA_TT

     S58 in S82 --- [in progress]

• Proteins with amber mutations
• T7 amber_TT

     S80 : in pSB1AK3

• GFP amber

     [in progress : transformation]

• GFP_TT

     [in progress : transformation]

• other constructs
• pT7_RBS_GFP_TT  :

     S79 :  this construct in ampR plasmid (pSB1AK3?).
     *Functionality has been tested through transformation in BL21 and microscopy. (cf.27/08/11)

To Do List

in order of priority:

Sequencing

• S92 = S81 in pHM3

Cloning

• The plates labelled with orange tape: "Danny's Transfos": c.f. 03/09/11

     In case of failure, redo the ligations the transform.
     In case of success, S38 in S24 has to be moved into pSB1C3. The others are already in this plasmid. They are to be inserted into pHM3, then transformed into subtilis.