The Expression Of Recombinant α-Cobratoxin In Yeast Pichia Pastoris

a-Cobratoxin, a kind of basic peptide from King Cobra venom with 72 amino acid residues and "three-finger" dimensional conformation, is regarded to possess affinity interaction with nicotinic acetylcholine receptor (nAChR), and also widely employed to investigate the structure and function of AChR. Studies show that a-Cobratoxin provides the important medical application of abirritation, restraining tumour and abstaining heroin. But the source of the product is restricted seriously because the King Cobra is the country's second grade protection animals. So gene engineering method is used to express the product in our experiment to deal with the problem.In the study, Yeast Pichia pastoris expression system, which was widely used in expressing the recombinant protein, was chosen as the gene expression system. a-Cobratoxin DNA was synthesized and inserted into the plasmid pPIC9K to construct recombinant plasmid pPIC9K/hPK followed by linearization with restricted endonuclease BglII. The subsequent plasmid was transformed into the host Pichia pastoris GS115 by eletroporation to form the transformant, which was incubated on MD/MM...