Reporter galactosidase

CL reaction can be catalyzed by enzymes other than HRP (e.g., microperoxidase and catalase) and by other substances [hemoglobin, cytochrome c, Fe(III), and other metal complexes]. The presence of suitable molecules such as phenols (p-iodophenol), naphthols (l-bromo-2-naphthol), or amines (p-anisidine) increases the light production deriving from the HRP-catalyzed oxidation of luminol and produces glow-type kinetics [6, 7], The use of other enzymes, such as glucose-6-phosphate dehydrogenase [38-41], P-galactosidase [42], and xanthine oxidase [43-46], as CL labels has been reported. 

The initial approach adopted entailed administration of naked plasmid DNA housing the gene of interest. This avenue of research was first opened in 1990, when it was shown that naked plasmid DNA was expressed in mice muscle cells subsequent to its i.m. injection. The plasmid DNA concerned housed the P-galactosidase gene as a reporter. Subsequent expression of P-galactosi-dase activity could persist for anything from a few months to the remainder of the animal s life. The transfection rate recorded was low (1-2 per cent of muscle fibres assimilated the DNA), and the DNA was not integrated into the host cell s chromosomes. 

There are other reporter gene systems, such as j8-galactosidase (a bacterial enzyme), chloramphenicol acetyltransferase (a bacterial enzyme), and aequorin (a jellyfish protein). 

Screen for colony growth by transcriptional selection Screen for p-galactosidase reporter gene activity... 

Thus, to summarize and to show that the two proteins interact, AH109 yeast cells transformed with the appropriate DNA-BD and AD vectors grow in -W, -L SD media and also in -W, -L, -H, -Ade SD media and yield a-galactosidase and p-galactosidase reporter gene activities. 

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