Part:BBa_K2762000

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 inserted the part on pSB1C3 and transformed the plasmid into DH5 alpha. After the the formation of the colony, we extracted the plasmid and screened the construction by enzyme digestion.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Unknown
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 1149
  Illegal AgeI site found at 252
  
• 1000
  COMPATIBLE WITH RFC[1000]

Source

Codon oprimized Synechococcus elongatus PCC7002.

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