Difference between revisions of "Part:BBa K2762000"

Revision as of 12:17, 14 October 2018

rbcL (Large subunit of the ribulose-bisphosphate carboxylase/oxygenase)

Background

Ribulose-1,5-biphosphate carboxylase/oxygenase catalyzes the first reaction of the Calvin cycle, converting the combination of Ribulose-1,5-biphosphate(RuBP) and carbon dioxide to two 3-phosphoglycerate molecular. The rbcL part encodes the large subunit of the RubisCo enzyme, which also contain the active site of the enzyme. In our carbon fixing pathway, the RubisCO enzyme is the most important enzyme catalyzing the reaction of RuBP and CO₂.

Mechanism

The rbcL gene is from cynobacteria Synechococcus elongatus PCC7002.The activation of RubisCO will be maximize with the binding of the RbcS subunit. To get more information, see our composite part: RubisCO

Expression in E. coli

We did two experiment to confirm the expression of this part. First we inserted the part on pSB1C3 and transformed the plasmid into DH5 alpha. We extracted the plasmid after the the formation of the colony and screened the construction by enzyme digestion. Second, we cloned the plasmid into BL21(DE3) and induced the promoter with IPTG. We then did the SDS-PAGE to confirm the present of the RbcL.

Sequence and Features