Fl-Galactosidase

Monitoring protein interactions by intracistronic fl-galactosidase complementation... 

The fl-galactosidase complementation assay has also been adapted for use in mammalian cells (Rossi et al., 1997). The availability of fluorescent substrates for (3-galactosidase allows for fluorescence microscopy and FACS analysis of mammalian cells expressing the fusion proteins of interest. Therefore, similar to the mDHFR system, fl-galactosidase complementation assays may prove useful for genome-scale studies of protein-protein interactions in mammalian cells. 

Rossi, F. M. V., Blakely, B. T., and Blau, H. M. (2000). Interaction blues protein interactions monitored in live mammalian cells by fl-galactosidase complementation. Trends Cell Biol. 10, 119-122. 

Recently the 0X26 immunohposomes were used in a gene dehvery approach to transport expression vectors for ludferase or fl-galactosidase through the BBB [116]. The plasmids... 

Lactose Lactase, fl-galactosidase Milk Products destined to lactose intolerant persons Cow and milk diagnosis... 

Tritylagarose (29) binds enzymes in a reversible fashion through hydrophobic forces, providing another kind of reversible immobilization. Several enzymes, such as alkaline phosphatase, fl-galactosidase, lactate dehydrogenase, and spleen phosphodiesterase, have been immobilized (29). 

Fig. 24.1 Simplified diagram of the plasmid pUCl 8. lacZ represents the insertional inactivation marker coding for fl-galactosidase activity. A multiple cloning site (MCS) is present within the LacZ gene to enable the cloning of DNA fragments. Ori represents the origin of replication which, in this case, works in Escherichia coli. Finally, Ampr represents an ampicillin resistance marker.

Koyama, Y, Okamoto, S., and Furukawa, K. (1990) Cloning of a-and fl-galactosidase genes from an extreme thermophile, Ihermus strain T2, and their expression in Ihermus thermophilus HB27. Appl Environ. Microbiol, 56, 2251—2254. 

Immobilized enzymes are often more stable than the soluble counterparts and can be recycled. They are also removable from a reaction mixture. PHA synthase fusions with the enzyme of interest were successfully developed and enabled production of recombinant enzyme already attached to a support material (PHA) in one step. The fl-galactosidase LacZ [21] was the first immobilized enzyme followed by alpha-amylase, organophosphohydrolase, and enzymes involved in sialic acid synthesis [76-78]. These demonstrated applicability PHA-immobilized enzymes in food processing, biomass conversion, bioremediation, and fine chemical synthesis. 

Fig.2. Glycosaminoglycans (mucopolysaccharides) and sites of attack by degradative enzymes. Absence or defective function of any one of these enzymes leads to accumulation of incompletely degraded mucopolysaccharide, i.e. mucopolysaccharidosis. 1. p-Glucuronidase 2. IV-Acetylgalactosamine-4-sulfatase 3. p-W-Acetylhexosaminidase 4. fl Acetylgalactosamine-6-sulfatase 5. p-Galactosidase 6. W-Acetylglucosamine-6-sulfatase 7. lduronate-2-sulfatase 8. L-lduronidase 9. o-A/-Acetylglucosami-nidase 10. Heparan Af-sulfatase.

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