Difference between revisions of "Part:BBa K424017:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | We were able to ligate successfully the promotor with the RBS into a plasmid backbone | + | We were able to ligate successfully the promotor with the RBS into a plasmid backbone and the reporter with the terminator; and we get the rhamnosiltransferase 1 gene enzyme mutated to be compatible with the Assembly Standard 10. We need to ligate all the parts to test our plataform for rhamnosyltransferase expression. |
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===Source=== | ===Source=== | ||
Revision as of 23:58, 27 October 2010
Test plataform for rhamnosyltransferase BioBrick (Rh1AB_BB) expression in E. coli
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 220
Illegal BamHI site found at 780
Illegal XhoI site found at 956
Illegal XhoI site found at 2242 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1135
Illegal NgoMIV site found at 1856
Illegal NgoMIV site found at 1969 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 445
Illegal BsaI site found at 1485
Illegal BsaI.rc site found at 629
Illegal BsaI.rc site found at 3135
Design Notes
We were able to ligate successfully the promotor with the RBS into a plasmid backbone and the reporter with the terminator; and we get the rhamnosiltransferase 1 gene enzyme mutated to be compatible with the Assembly Standard 10. We need to ligate all the parts to test our plataform for rhamnosyltransferase expression.
Source
iGEM, Part registry organization give us the composite parts. The standard Rh1AB part was gene isolated from a Pseudomonas aeruginosa and mutated by a mutagenesis protocol.
References
Assembly Standard Protocol 10, www.parts.igem.org
QuikChange Lightning Multi Site-Directed Mutagenesis Kit Instruction manual, catalog #210515 from Stratagene.