Difference between revisions of "Part:BBa K2127001"

Revision as of 14:38, 21 October 2016

Inducible High Expression His-Tagged Photosystem II CP47 subunit

Cyanobacteria is an not optimal system to manipulate the protein expression mainly due of the lack of strong promoters. Pcpc560 is a Cyanobacterial super promoter discovered by Dr. Ma’s team that contains two predicted promoter sites and a transcription binding site containing 14 homologous bacterial regions upstream of the regulated cpcB gene. Using Pcpc560, Dr. Ma’s team demonstrated that functional protein was produced at 15% of the total cell protein volume. Our team decided to test the effect of the transcription factor binding site on protein production in E.Coli cells DH5α. We believe that the predicted 14 homologous binding sites from Pcpc560 will allow us to increase the protein production greatly using the E.Coli DH5α. We constructed a biobrick containing the 14 transcription binding sites followed by a strong RBS (BBa_B0030) and a mutant HIS-Tag psbB gene. The super promoter can be an valuable asset to other iGEM teams for protein production and therefore we have decided to submit the Construct along with the Transcription Binding Site from Pcpc560.

We envision that this construct will be helpful in allowing teams to directly begin working with Photosystem components. Being sent as an expressible construct, teams will have the option of either using the construct directly for protein work, or isolating the genetic sequence and manipulating it to their needs.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
COMPATIBLE WITH RFC[12]
21
INCOMPATIBLE WITH RFC[21]
Illegal BamHI site found at 658
23
COMPATIBLE WITH RFC[23]
25
INCOMPATIBLE WITH RFC[25]
Illegal AgeI site found at 782
Illegal AgeI site found at 1379
Illegal AgeI site found at 1583
1000
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI.rc site found at 1103
Illegal SapI site found at 1780

@@ Line 5: / Line 5: @@
 Cyanobacteria is an not optimal system to manipulate the protein expression mainly due of the lack of strong promoters. Pcpc560 is a Cyanobacterial super promoter discovered by Dr. Ma’s team that contains two predicted promoter sites and a transcription binding site containing 14 homologous bacterial regions upstream of the regulated cpcB gene. Using Pcpc560, Dr. Ma’s team demonstrated that functional protein was produced at 15% of the total cell protein volume. Our team decided to test the effect of the transcription factor binding site on protein production in E.Coli cells DH5α. We believe that the predicted 14 homologous binding sites from Pcpc560 will allow us to increase the protein production greatly using the E.Coli DH5α. We constructed a biobrick containing the 14 transcription binding sites followed by a strong RBS (BBa_B0030) and a mutant HIS-Tag psbB gene. The super promoter can be an valuable asset to other iGEM teams for protein production and therefore we have decided to submit the Construct along with the Transcription Binding Site from Pcpc560.
-We envision that this construct will be helpful in allowing teams to directly begin working with Photossytem components. Being sent as an expressible construct, teams will have the option of either using the construct directly for protein work, or isolating the genetic sequence and manipulating it to their needs.
+We envision that this construct will be helpful in allowing teams to directly begin working with Photosystem components. Being sent as an expressible construct, teams will have the option of either using the construct directly for protein work, or isolating the genetic sequence and manipulating it to their needs.
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