This part contains the reverse complementary of a fragment of E5 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 E5 from HPV-16, it interacts with the riboswitch, allowing the expression of the attached coding sequence.
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 4: BBa_K1905000 - E5 Riboswitch ligation
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Revision as of 18:28, 24 October 2016
Riboswitch to detect E5 mRNA from HPV-16
DNA coding for a riboswitch to detect E5 mRNA from Human Papillomavirus type 16. This part includes a promoter (BB_R0010) and a RBS (BBa_B0034), the reverse complementary for a section of E5 DNA coding region, and the original sequence with one base changed. When there is presence of E5 mRNA, the riboswitch turns on and it allows the expression of the reporter protein.
