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

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.