LacZ Reporter Gene (Restriction Cloning Compatible)
LacZ is a reporter gene that encodes the beta-galactosidase, an enzyme commonly used for the blue-white technique. Bacteria encoding the LacZ gene, and thereby producing beta-galactosidase, cultured in the presence of Xgal will produce a prominent blue color. In contrast with other reporter proteins such as Green Fluorescent Protein which may not always appear visible without fluorescent screening, LacZ-encoding bacteria produce a powerful blue color demonstrating successful color without any additional color screening. Through this, LacZ is an efficient reporter, and we have chosen to incorporate it in our toehold switch design.
Toehold switches are RNA-based riboregulators functioning as switches that can activate/inhibit reporter expression. Using a complementary trigger sequence, reporter expression can be activated through the binding of the trigger sequence to the toehold switch, resulting in the uncoupling of the secondary structure, allowing the RBS to be freed from the "closed" position, allowing for translation of the reporter protein.
This part encodes the full-length LacZ Beta-galactosidase gene with a wobble introduced to ensure that the gene is restriction fragment cloning compatible. In our model, LacZ is the downstream reporter translated following binding of the trigger sequence to the toehold switch.
Sequence and Features
- 10COMPATIBLE WITH RFC
- 12COMPATIBLE WITH RFC
- 21COMPATIBLE WITH RFC
- 23COMPATIBLE WITH RFC
- 25COMPATIBLE WITH RFC
- 1000COMPATIBLE WITH RFC