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Gene Synthesis> | His-tagged nChiAvd [17] was produced in BL21-AI cells using pYU25 plasmid (gift from Prof. Lloyd Ruddock, University of Oulu) as described by Hytönen et al. [34]. pYU25 encodes a polycistronic cassette containing N-terminally His-tagged nChiAvd with the components for cytoplasmic disulfide bond formation (ERv1p and PDI). It is similar in structure to pKEHS1160 [35] which encodes BMP4 plus the components required for cytoplasmic disulfide bond formation, except that in pYU25, the ERv1p and PDI codons are optimized for E. coli expression and the T7 promoter has been replaced with a Ptac promoter. Codon optimization and gene synthesis were performed by GenScript. | Get A Quote |
In biosensor experiments, ligand molecules need to be immobilized tightly on sensor surfaces for sensitive and robust detection of the analyte of interest. Stretches of histidines (His-tags) are typically inserted into recombinant proteins for affinity purification purposes, but can also be used in the immobilization of biosensors functionalized with Ni-NTA (nitrilotriacetic acid). The His-tag-Ni-NTA bond is, however, easily disrupted by factors including low pH, reducing agents, or chelating agents such as EDTA. In biosensing applications, a stable ligand immobilization that tolerates harsh conditions is preferable, because the sample matrix can vary from bodily fluids to waste waters. We describe here Co(III)... More