Part:BBa_K1905001
Riboswitch to detect L1 mRNA from HPV
DNA coding for a riboswitch to detect L1 mRNA from Human Papillomavirus. This part includes a promoter (BB_R0010) and a RBS (BBa_B0034), the reverse complementary for a section of L1 DNA coding region, and the original sequence with one base changed. When there is presence of L1 mRNA, the riboswitch turns on and it allows the expression of a reporter 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]
This part contains the reverse complementary of a fragment of L1 sequence from HPV-16, the original with a base changed, RBS and promoter (for overexpress) . After translation it folds in a shape that blocks the access of the ribosome to the RBS, so an attached protein coding sequence will not express. When there is presence of mRNA L1 from HPV-16 and possibly from other HPVs or viruses, it interacts with the riboswitch, allowing the expression of the attached coding sequence.
Figure 1. Biobrick design diagram
This biobrick was chemically synthesised and cloned into both pSB1A3 and pSB1C3 vectors. These were transformed into E. coli TOP10 strain. It was grown at 37°C for 12 hours and then propagated on liquid LB medium. Miniprep plasmid extraction was carried out in order to document the plasmid, as observed on Figure 2.
Figure 2. Agarose (0.8%) gel electrophoresis: Miniprep plasmid extraction. Lane 7: BBa_K1905001 - L1 Riboswitch ligation
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