Journal:

Article Title: PDI Improves Secretion of Redox-Inactive β-glucosidase

doi: 10.1021/bp060287p

Figure Lengend Snippet: Comparison of relative PDI and intracellular β-glucosidase levels during β-glucosidase expression in different yeast strains after exponential growth phase (48 h) in batch culture normalized to wild-type β-glucosidase in BJ5464. 1 OD-mL yeast cells expressing β-glucosidase in 25 mL batch culture were collected and lysed as described in Materials and Methods. Cell lysates were subjected to SDS-PAGE and the intracellular PDI was detected by Western blotting. The PDI levels relative to the parent strain BJ5464+pRS314-β were determined using ImageQuant software (Molecular Dynamics). The range of PDI levels is based on five sets of Western blot data.

Article Snippet: Plasmids The single copy expression plasmid pRS314-β was constructed from a secretion-modified full-length β-glucosidase with an N-terminal synthetic pre-pro secretion signal sequence and a 10-amino-acid C-terminal c- myc detection tag under the control of a GAL1-10 promoter and the plasmid pRS314 (ATCC) containing the TRP1 marker as reported previously 5 .

Techniques: Comparison, Expressing, Western Blot, Software