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Updated: Jul 19, 2026
High Throughput Quantitative Expression Screening and Purification Applied to Recombinant Disulfide-rich Venom Proteins Produced in E. coli
Published on: July 30, 2014
Soluble expression, purification and functional identification of a disulfide-rich conotoxin derived from Conus
Canhui Pi1, Junliang Liu, Lei Wang
1State Key Laboratory of Biocontrol, Guangdong Province Key Laboratory of Therapeutic Functional Genes, Open Laboratory for Marine Functional Genomics, Department of Biochemistry, College of Life Sciences, Sun Yat-sen University, People's Republic of China.
Abstract:
Conotoxins are a diverse array of small peptides mostly with multiple disulfide bridges. These peptides become an increasing significant source of neuro-pharmacological probes and drugs as a result of the high selectivity for ion channels and receptors. Usually, the analogue of natural conotoxins is produced by means of chemical synthesis. Here, we present a simple and fast strategy of producing disulfide-rich conotoxins via recombinant expression. By fused with thioredoxin and His tag, a novel O-superfamily conotoxin lt7a was successfully expressed in Escherichia coli and purified, resulting in a high yield of recombinant lt7a about 6 mg/l. The purity of target protein is up to 95% as identified by HPLC results. Whole cell patch-clamp recording revealed that the new conotoxin blocked voltage-sensitive sodium channels in rat dorsal root ganglion neurons, indicating it might be a novel microO-conotoxin.
