Difference between revisions of "Part:BBa K5115053"
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===Introduction=== | ===Introduction=== | ||
| − | This composite part combines all the subunits, except hoxU, of the hydrogenase in our ribozyme-assisted polycistronic co-expression system：pRAP. Actually, compared with [https://parts.igem.org/Part:BBa_K5115052 ribozyme connected hox and hyp, without hoxF], there is only one difference: one deleted hoxF and another deleted hoxU. The deleted subunits are all used to link with EP. After comparison between experiments of these two designs, we make sure that the hoxF-linking-EP one is more satisfying. So, to learn more about the | + | This composite part combines all the subunits, except hoxU, of the hydrogenase in our ribozyme-assisted polycistronic co-expression system：pRAP. Actually, compared with [https://parts.igem.org/Part:BBa_K5115052 ribozyme connected hox and hyp, without hoxF], there is only one difference: one deleted hoxF and another deleted hoxU. The deleted subunits are all used to link with EP. EP ligation of different subunits leads to different effects of hydrogenase's entry into the carboxysome, which further affects the effect of our overall design. After comparison between experiments of these two designs, we make sure that the hoxF-linking-EP one is more satisfying. So, to learn more about the final adopted part, please click on [https://parts.igem.org/Part:BBa_K5115052 ribozyme connected hox and hyp, without hoxF]. |
===Characterization=== | ===Characterization=== | ||
Revision as of 14:39, 1 October 2024
ribozyme connected hox and hyp, without hoxU
Introduction
This composite part combines all the subunits, except hoxU, of the hydrogenase in our ribozyme-assisted polycistronic co-expression system：pRAP. Actually, compared with ribozyme connected hox and hyp, without hoxF, there is only one difference: one deleted hoxF and another deleted hoxU. The deleted subunits are all used to link with EP. EP ligation of different subunits leads to different effects of hydrogenase's entry into the carboxysome, which further affects the effect of our overall design. After comparison between experiments of these two designs, we make sure that the hoxF-linking-EP one is more satisfying. So, to learn more about the final adopted part, please click on ribozyme connected hox and hyp, without hoxF.
Characterization
Agarose gel electrophoresis
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| Figure 1. Agarose gel electrophoresis of PCR products amplified from one E. coli (DH5α) colony.
M: DNA Marker. Lanes 1,3-8: Amplification of specific regions corresponding to hoxF, hoxY, hoxH, hoxW, hoxI, hypA, and hypB, demonstrating successful assembly and integrity of the ribozyme-connected hox and hyp, without hoxU (no band in lane 2) as designed. Primers for these PCR are listed on https://2024.igem.org/fudan/parts. |
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 654
Illegal BglII site found at 1472
Illegal BglII site found at 1765
Illegal BamHI site found at 2059
Illegal XhoI site found at 99
Illegal XhoI site found at 2266
Illegal XhoI site found at 4505
Illegal XhoI site found at 6334
Illegal XhoI site found at 7496
Illegal XhoI site found at 9335 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 533
Illegal NgoMIV site found at 1071
Illegal NgoMIV site found at 1281
Illegal NgoMIV site found at 1593
Illegal NgoMIV site found at 2359
Illegal NgoMIV site found at 3039
Illegal NgoMIV site found at 5646
Illegal AgeI site found at 6821 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 4159
Illegal BsaI.rc site found at 702
Illegal BsaI.rc site found at 1206