title: Synthesis, expression and purification of a type of chlorotoxin-like peptide from the scorpion, Buthus martensii Karsch, and its acute toxicity analysis.
abstract: A gene, rBmK Cta, encoding a chlorotoxin-like peptide from the scorpion, Buthus martensii Karsch, was synthesized according to the sequence optimized for codon usage in Escherichia coli and was expressed in E. coli BL21 (DE3) using a pExSecI expression system in which the IgG-binding domain-ZZ of protein A is fused to the N-terminal of rBmK CTa. The fusion protein, ZZ-rBmK CTa, was expressed in soluble form (7.8 mg l(-1)) and was purified to give a single band on SDS-PAGE. The domain-ZZ of fusion protein ZZ-rBmK CTa was removed by cleavage of an Asn-Gly peptide bond with hydroxylamine. The rBmK CTa was separated from the IgG-binding moiety by a second passage through the IgG affinity column. Western blot analysis demonstrated that this protein was rBmK CTa. Acute toxicity assay in mice demonstrated that the rBmK CTa had an LD(50) value of 4.3 mg kg(-1).
authors.affiliation: Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, 030006, Taiyuan, People's Republic of China. aliang@sxu.edu.cn
authors.forename: Yue-jun
authors.initials: YJ
authors.lastname: Fu
authors.forename: Li-tian
authors.initials: LT
authors.lastname: Yin
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Bao-feng
authors.initials: BF
authors.lastname: Chai
authors.forename: Ai-hua
authors.initials: AH
authors.lastname: Liang
mesh_terms: D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D015870:Gene Expression; D008297:Male; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000633:toxicity; D012604:Scorpion Venoms / Q000096:biosynthesis* / Q000302:isolation & purification / Q000633:toxicity*; D012605:Scorpions / Q000235:genetics*; D013205:Staphylococcal Protein A / Q000096:biosynthesis / Q000302:isolation & purification / Q000633:toxicity
chemical_list: C516430:BmK CTa protein, Buthus martensii; D011993:Recombinant Fusion Proteins; D012604:Scorpion Venoms; D013205:Staphylococcal Protein A; C091539:Chlorotoxin