Difference between revisions of "Part:BBa K2762002"
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===Mechanism=== | ===Mechanism=== | ||
| − | The rbcS genes are from cyanobacteria Synechococcus elongatus PCC7002. We designed the T7 promoter (BBa_I719005)for the gene and cloned gene in BL21 to ensure the high expression rate. It has reported that the expression of RubisCO could not be detectable while only add single promoter of the rbcL-rbcX-rbcS gene fragment. Therefore we added another T7 promoter on the upstream of rbcX-rbcS gene and named the part as rbcXS. We also did the coden optimization for the gene to ensure successful expression. The activation of RubisCo will be maximize after the binding of the RbcS subunit. To get more information, see our composite part: RubisCO. | + | The rbcS genes are from cyanobacteria <i>Synechococcus elongatus</i> PCC7002. We designed the T7 promoter (BBa_I719005)for the gene and cloned gene in BL21 to ensure the high expression rate. It has reported that the expression of RubisCO could not be detectable while only add single promoter of the rbcL-rbcX-rbcS gene fragment. Therefore we added another T7 promoter on the upstream of rbcX-rbcS gene and named the part as rbcXS. We also did the coden optimization for the gene to ensure successful expression. The activation of RubisCo will be maximize after the binding of the RbcS subunit. To get more information, see our composite part: RubisCO. |
===Expression in <i>E. coli</i>=== | ===Expression in <i>E. coli</i>=== | ||
Revision as of 10:11, 16 October 2018
Small subunit of the ribulose-bisphosphate carboxylase/oxygenase rbcS
Backgriund
The rbcS part encode the rbcS subunit of RubisCO. The rbcS affect the activity of RubisCO.
Mechanism
The rbcS genes are from cyanobacteria Synechococcus elongatus PCC7002. We designed the T7 promoter (BBa_I719005)for the gene and cloned gene in BL21 to ensure the high expression rate. It has reported that the expression of RubisCO could not be detectable while only add single promoter of the rbcL-rbcX-rbcS gene fragment. Therefore we added another T7 promoter on the upstream of rbcX-rbcS gene and named the part as rbcXS. We also did the coden optimization for the gene to ensure successful expression. The activation of RubisCo will be maximize after 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 cloned the plasmid into DH5 alpha. We extracted the plasmid after the colony formed and did the enzyme digestion to confirm the insertion was successful. Second, we cloned the plasmid into BL21(DE3) and induced the promoter with IPTG. We than did the SDS PAGE to confirm the present of the RbcX and RbcS protein.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]