Difference between revisions of "Part:BBa K1041000"
(→Characterisation) |
|||
| Line 2: | Line 2: | ||
<partinfo>BBa_K1041000 short</partinfo> | <partinfo>BBa_K1041000 short</partinfo> | ||
| − | + | Team NRP-UEA (2013) used mutagenesis to add a Nde1 site at the start of the RFP coding region of the biobrick BBa_J04450. This enables a restriction digest with Nde1 to be performed allowing the RFP coding region to be excised from the plasmid. | |
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
| Line 19: | Line 19: | ||
==Characterisation== | ==Characterisation== | ||
| − | + | Characterisation of this biobrick involved comparisons with the original Bba_J04450 biobrick by performing transformations, restriction digests and BLAST analysis. | |
| + | ===Transformation=== | ||
| + | |||
| + | Parts BBa_K1041000 and BBa_J04450 were transformed into Alpha-Select competent ''E.Coli'' cells and plated onto LB Agar plates which contained have colonies that appear red under natural light ''fig.1''. The addition of a NdeI site has not effected the ability for the RFP gene to be transcribed. | ||
[[Image:WP 000051.jpg|thumb|Fig 1:(Left) BBa_K1041000 and (right) BBa_J04450 visualized under non-UV lightbox]] | [[Image:WP 000051.jpg|thumb|Fig 1:(Left) BBa_K1041000 and (right) BBa_J04450 visualized under non-UV lightbox]] | ||
Revision as of 12:27, 6 September 2013
RFP Coding Device
Team NRP-UEA (2013) used mutagenesis to add a Nde1 site at the start of the RFP coding region of the biobrick BBa_J04450. This enables a restriction digest with Nde1 to be performed allowing the RFP coding region to be excised from the plasmid.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 781
Illegal AgeI site found at 893 - 1000COMPATIBLE WITH RFC[1000]
Composite part of the following BioBricks:
| LacI R0010 | B0034 | mRFP1 E1010 | B0015 | |
 |  |  |  |
- BBa_R0010: Promoter (lacI regulated)
- BBa_B0034: RBS (Elowitz 1999)
- BBa_E1010: Red Fluorescent Protein from Discosoma striata
- BBa_B0015: Double Terminator
Characterisation
Characterisation of this biobrick involved comparisons with the original Bba_J04450 biobrick by performing transformations, restriction digests and BLAST analysis.
Transformation
Parts BBa_K1041000 and BBa_J04450 were transformed into Alpha-Select competent E.Coli cells and plated onto LB Agar plates which contained have colonies that appear red under natural light fig.1. The addition of a NdeI site has not effected the ability for the RFP gene to be transcribed.
