Part:BBa_K4140024

PAH releasing device

Part Description

The LacZ-alpha fragment is encoded by this region and is derived from the pUC19 cloning vector. Because it is smaller than the LacZ-omega region, the LacZ-alpha fragment can be easily incorporated into a plasmid when the two non-functional LacZ gene fragments (alpha and omega) are co-expressed. X-gal (5-bromo-4-chloro-3-indoyl-d-galactopyranoside), a soluble colourless molecule that is a substrate of ß-galactosidase and generates a blue product when cleaved, is used to detect the alpha-complementation. It's linked to KP-SP ISA secreting peptide signal at the N-terminus of the reporter protein to transmit it from intra cellular to extra cellular

Usage

In order to detect the increased levels of phenylalanine in PKU patients, we used the lacZ alpha gene which encodes beta-galactosidase, which catalyzes the cleavage of lactose to form galactose and glucose that utilizes X-gal to change the color into blue as shown in figure 1.

Figure 1. (shows the whole cell-based biosensor.)

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 383
  Illegal BamHI site found at 910
  Illegal XhoI site found at 620
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]