Difference between revisions of "Part:BBa K2762005"

Revision as of 13:19, 17 October 2018

P_T7-B0034-rbcL-B0015

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. We designed the T7 promoter (BBa_I719005) for the gene and cloned gene in BL21 (DE3) to ensure the high expression rate because the RbcL protein is the most important protein as previous description. We also coden optimized the gene to ensure successful expression. However, the activation of RubisCO will be maximize with the binding of the RbcS subunit. To get more information, see our composite part: BBa_K2762011.

Characterization

Expression in E. coli

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 consruction by enzyme digestion.

Sequence and Features

References

[1] Janet Newman 1t and Steven Gutteridge2*. (1994,JUNE.15). Structure of an effector-induced inactivated state of ribulose 1,5-bisphosphate carboxylase/oxygenase: the binary complex between enzyme and xylulose 1,5-bisphosphate. cell. Doi:https://doi.org/10.1016/S0969-2126(00)00050-2