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Overview

We have built 5 new Basic Parts and 5 Composite Parts in total. Five Composite Parts are used to express CA,CA2,PRK,CA-PRK and CA2-PRK respectively.

Basic Parts

Description Type Name
CYC1 Terminator BBa_K3812006
BW25113 RBS BBa_K3812011
6xHis-tag Tag BBa_K3812012
Paox1 Promoter BBa_K3812005
pSB1C3 Plasmid BBa_K3812014
CA2 Coding BBa_K4112005
CA Coding BBa_K4112006
PRK Coding BBa_K4112007
CA2-PRK Coding BBa_K4112008
CA-PRK Coding BBa_K4112009

Composite Parts

Description Type Name
CA2 Composite BBa_K4112010
CA Composite BBa_K4112011
PRK Composite BBa_K4112012
CA2-PRK Composite BBa_K4112013
CA-PRK Composite BBa_K4112014

Validation

The PTCA-encoded carbonic anhydrase of P. tricornutum was cloned by overlap PCR. As a member of the β-CA family, it can reversibly catalyze the CO2 hydration reaction, which can improve CO2 conversion efficiency and play a key role in CCM, as well as in ion exchange, CO2 acquisition, and photosynthesis.

PTCA2, which is highly homologous to PTCA, encodes a protein with fully conserved β-CA zinc coordination residues and significant levels of mRNA accumulation at low CO2 concentrations.

PRK encodes phospho-ribulose kinase, an enzyme specific to the Calvin cycle, which plays a crucial role in regulating the flow of sugars in the Calvin cycle by catalyzing the formation of the receptor ribulose-1,5-bisphosphate of CO2 with the help of ATP phosphorylation.

Use linker to connect them separately, build CA-PRK and CA2-PRK, transferred to E. coli and tested.

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