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Proteins, Expression, Isolation and Analysis | … Page 2. 846 Kim et al. chemically synthesized (GENSCRIPT, USA; Fig. 1B). The synthesized DNA fragment was digested and ligated into the NcoI and XhoI sites of pProEX HTa (Novagen), which contains an N- terminal hexahistidine tag … | Get A Quote |
TolC is an outer membrane porin protein and an essential component of drug efflux and type-I secretion systems in Gram-negative bacteria. TolC comprises a periplasmic alpha- helical barrel domain and a membrane-embedded beta-barrel domain. TdeA, a functional and structural homolog of TolC, is required for toxin and drug export in the pathogenic oral bacterium Actinobacillus actinomycetemcomitans. Here, we report the expression of the periplasmic domain of TdeA as a soluble protein by substitution of the membraneembedded domain with short linkers, which enabled us to purify the protein in the absence of detergent. We confirmed the structural integrity of the TdeA periplasmic domain by size-exclusion chro... More