Chemical chaperone therapy: Luciferase assay for screening of β-galactosidase mutations

Isolation and Screening of Fungal Strains for β-Galactosidase Production

Enzymes are the biocatalysts which catalyze the biochemical processes and thus have a wide variety of applications in the industrial sector. β-Galactosidase (E.C. 3.2.1.23) also known as lactase, is one of the prime enzymes, which has significant potential in the dairy and food processing industries. It has the capability to catalyze both the hydrolytic reaction for the production of lactose hy...

An In Vitro Assay of β-Galactosidase from Yeast

mouse genotyping by the three-primer multiple PCR. However, both of the smaller PCR products of 1068 (specific for the wild-type allele of mGluR4) and 1170 bp (specific for the mGluR4 transgene) were visible. Thus, all three allele combinations were clearly distinguishable on agarose gels stained with ethidium bromide. DNA from wildtype mice give a single smaller band (1068 bp; see Figure 2B, l...

LightSwitchTM Luciferase Assay System

Microplate Diffusion Assay for Screening of β-Glucanase-Producing Microorganisms

A method is described to screen fungal strains rapidly for overexpression of extracellular β-1,4-endoglucanase in the presence of high levels of sugar compounds. The semi-quantitative assay utilizes microplates in a 96-well format and an azurine dye covalently cross-linked (AZCL) chromogenic substrate. The digestion of AZCL-hydroxyethyl-β-1,4-endoglucanase results in the release of a blue dye d...

Development of a high-throughput screening cancer cell-based luciferase refolding assay for identifying Hsp90 inhibitors.

The 90 kDa heat-shock protein (Hsp90) and other cochaperones allow for proper folding of nascent or misfolded polypeptides. Cancer cells exploit these chaperones by maintaining the stability of mutated and misfolded oncoproteins and allowing them to evade proteosomal degradation. Inhibiting Hsp90 is an attractive strategy for cancer therapy, as the concomitant degradation of multiple oncoprotei...