IGEM:MIT/2006/Autotrophic E.coli/Biobrick Calvin cycle enzymes

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Background

  • prkA contains a Pst1 site
  • rbcL contains an EcoR1 site

Procedure

  • Miniprep AE4 (this will provide a plasmid prep of both enzymes).
  • Site-directed mutagenesis of the prkA and rbsLS
  • Sequence verify that we got mutations
  • Cut out the coding regions with primers containing BB ends.
  • Clone into BB vector.
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