Products/Services Used | Details | Operation |
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Proteins, Expression, Isolation and Analysis | … Two ExpressPlus™ PAGE Gel (GenScript, Piscataway, NJ, USA) 4–20% polyacrylamide gradient gels loaded with samples along with PageRuller Unstained Protein Ladder (Thermo Fisher Scientific) for the SDS‐PAGE assay and Odyssey Protein Molecular Weight Marker (LI … | Get A Quote |
Cyclodextrin glycosyltransferase (CGTase) enzyme is used industrially to obtain cyclodextrins. In this study, a newly isolated alkaliphilic CGTase producing Bacillus sp. SD5 was used as the source organism for the gene encoding for CGTase. The gene was amplified and sequenced in preparation for cloning into Pichia pastoris for methanol‐induced expression and extracellular secretion. Three strategies were employed in designing the amplification primers for obtaining the fragments to be used in transformation. Hence, three P. pastoris X33 clones were obtained: N2 (with natural gene sequence), S3 (also encoding the c‐myc epitope and polyhistidine tail), and M5 (encoding the polyhistidine tail). Extracellular C... More