Sensing system of Vitamin B12 coupled with Ldh
This part is a composite part that can be divided into two units. The first unit is a Tet R protein (BBa_C0040) , which is regulated by a constitutive promoter (BBa_J23106) and a double terminator BBa_B0015 . However, the translation of the Tet R protein is also regulated by a Cobalamin Riboswitch (BBa_K3933003) , which sequesters the RBS (BBa_B0032) when bound to vitamin B12.
The second unit is the reporter system related to the sensing of Vitamin B12 realised in the first part. It is composed of a Tet Operator (BBa_R0040) , which hinders the expression of the coding sequence placed after when it is bound by Tet R. It is followed by a Lactate Dehydrogenase (BBa_K3102006) translated thanks to a RBS (BBa_B0032) . This unit is also closed by the double terminator BBa_B0015 .
When the riboswitch isn't bound to Cobalamin, the RBS is not sequestered and Tet R is expressed, which hinders the expression of the Lactate Dehydrogenase.
However, when Cobalamin (or vitamin B12) binds to the riboswitch, it sequesters the RBS, and the Tet R is not translated. With no Tet R binding the Tet Operator, RFP can be translated and quantified. Thus, the RFP can be detected when Cobalamin binds to the riboswitch, thanks to a double inhibition system.
Thanks to Biorender
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12Illegal NheI site found at 7
Illegal NheI site found at 30
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25Illegal AgeI site found at 1720
- 1000Illegal SapI.rc site found at 1450