Part:BBa_K4673027
lacZα is a common reporter gene within E. coli
The lacZα gene has been widely employed as a reporter gene for detecting gene expression patterns. It encodes for an enzyme called β-galactosidase that hydrolyzes disaccharide lactose into glucose and galactose and metabolizes the substrate X-gal that offers visible colors. In addition, lacZα sequence carries multiple replication endpoints(MCS). The production of β-galactosidase will be affected if the lacZα gene is inserted by foreign DNA.
As one of the important parts, the lacZα gene was engineered into the pGEM-T EASY vector, enabling us to confirm the accurate insertion of the Lumbrokinase gene using blue-white screening. In blue-white screening, the deletion mutant of the lacZ gene(lacZΔM15) in the host E. coli strain carries an ω-peptide while the foreign plasmid containing lacZα gene carries a β-galactosidase α-peptide. Neither of them is functional on their own. However, if the foreign plasmid is transferred into the host E.coli that has the lacZΔM15, the two peptide sequences are able to express together to produce functional β-galactosidase enzymes. On the contrary, functional β-galactosidase enzymes will not be formed if the MCS on the lacZα gene is occupied with foreign DNA.
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]
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