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| Codon Optimization | Previously, we circumvented this outcome using a bacterial strain, SMC4, specifically derived for this purpose (Chalfie et al., 2003; Goodman et al., 2002) Here, we used an alternative strategy that enables us to propagate expression plasmids in standard bacterial strains (NEB 5-alpha Competent E. coli, High Efficiency): synthetic cDNAs codon-optimized for expression in insect cells. Accordingly, we obtained plasmids containing synthesized codon-optimized cDNAs encoding full-length MEC-4, MEC-10 and DEGT-1 (GenScript) in the pGEM-HE oocyte expression vector (Liman et al., 1992). DEL-1 was codon-optimized for expression in C. elegans (IDT) based on the predicted sequence reported in wormbase release WS250. | Get A Quote |
The degenerin channels, epithelial sodium channels, and acid-sensing ion channels (DEG/ENaC/ASICs) play important roles in sensing mechanical stimuli, regulating salt homeostasis, or responding to acidification in the nervous system. They share a common topology with two transmembrane domains separated by a large extracellular domain and are believed to assemble as homomeric or heteromeric trimers into non-voltage gated, sodium-selective, and amiloride-sensitive ion channels. Amiloride is not the only drug that targets DEG/ENaC/ASICs, however, they are also emerging as a target of nonsteroidal anti-inflammatory drugs (NSAIDs) as well as other classes of small molecules. C. elegans has about 30 genes encoding ... More
