| Products/Services Used | Details | Operation |
|---|---|---|
| Gene Synthesis | New expression systems were designed and constructed with the NONPs using the determined putative promoter regions to drive the gene expression with Saccharomyces cerevisiae α-mating factor pre-pro sequences (α-MF) as secretion signal leader. The promoters were placed instead of PGAP in the parent plasmid pGAPZαA::hGH, having NsiI and EcoRI restriction sites in 5′ and 3′ ends (Fig. 1). The complete sequences of the putative promoter regions were synthesized by GenScript (Piscataway, NJ, USA). The synthesized DNA sequences and parent plasmid were double-digested with NsiI and EcoRI, and then ligated. The constructed plasmids were transformed into E. coli. The recombinant plasmids carrying novel promoters were isolated from the transformants and sequences were analysed (ABI Prism 310 Genetic Analyzer, Applied Biosystems, USA). The sequence authenticity was checked with NCBI (NIH, USA) nucleotide BLAST tool. | Get A Quote |
Novel strong promoter discovery is crucial for the design of novel strains of the industrial yeast Pichia pastoris for recombinant protein (r-protein) production. In order to remedy the deficiency, transcriptome and proteome data of P. pastoris were analysed. Genes having higher expression levels than glyceraldehyde-3-phosphate-dehydrogenase (GAP) gene were identified as promoter sources. Pyruvate kinase- (PPYK) and pyruvate decarboxylase- (PPDC) promoters around pyruvate-node were determined as promising candidates, and in silico analysis of the putative promoter regions was performed. The putative promoters were introduced into the plasmid pGAPZαA::hGH harboring human growth hormone (hGH) gene, instea... More
