Difference between revisions of "Part:BBa K1905002"

 
 
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<partinfo>BBa_K1905002 parameters</partinfo>
 
<partinfo>BBa_K1905002 parameters</partinfo>
 
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This part contains the reverse complementary of a fragment of E6 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 E6 from HPV-16 and possibly from HPV-18, it interacts with the riboswitch, allowing the expression of the attached coding sequence.
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https://static.igem.org/mediawiki/2016/7/7e/TecCEMHSRE6BP.png
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Figure 1. Biobrick design diagram
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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.
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https://static.igem.org/mediawiki/2016/2/22/TecCEMHS_documentation2016.jpeg
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Figure 2. Agarose (0.8%) gel electrophoresis; Miniprep plasmid extraction. Lane 6: BBa_K1905002 - E6 Riboswitch ligation

Latest revision as of 18:32, 24 October 2016


Riboswitch to detect E6 mRNA from HPV

DNA coding for a riboswitch to detect E6 mRNA from Human Papillomavirus type 16 and type 18. This part includes a promoter (BB_R0010) and a RBS (BBa_B0034), the reverse complementary for a section of E6 DNA coding region, and the original sequence with one base changed. When there is presence of E6 mRNA, the riboswitch turns on and it allows the expression of a reporter protein.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


This part contains the reverse complementary of a fragment of E6 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 E6 from HPV-16 and possibly from HPV-18, it interacts with the riboswitch, allowing the expression of the attached coding sequence.


TecCEMHSRE6BP.png


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.


TecCEMHS_documentation2016.jpeg


Figure 2. Agarose (0.8%) gel electrophoresis; Miniprep plasmid extraction. Lane 6: BBa_K1905002 - E6 Riboswitch ligation