C-type lectin superfamily in snake comprises a group of proteins which display anticoagulation or various other functions. PCR primers were designed according to N-terminal sequence of a snake venom protein of which the complete amino acid sequence had been unknown. Total RNA was extracted from venom gland of Agkistrodon acutus from Hunan Province, and was reversely transcribed to cDNA. Then by applying the touchdown method to anchored-PCR, with only one specific and one universal primer, two relatively distinct bands were amplified in a single non-nested PCR reaction of no more than 30 cycles. Cloning and sequencing indicated that the sequence is homologous with other members of the C-type lectin superfamily in snake. High level fusion expression in E.coli was achieved, with the fusion protein accounting for 26%～30% of total proteins.