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Abstract
Based on the known amino acid sequences of BmK AS and BmK AS-1, the gene specific
primers were designed and synthesized for 3' and 5' RACE (Rapid Amplification of cDNA
Ends). Their partial cDNA fragments obtained by 3' and 5' RACE were cloned and sequenced,
and the full length cDNA sequences of BmK AS and BmK AS-1 were then completed by overlapping
their two partial cDNA sequences, respectively. The predicted amino acid sequences
both consist of 85 amino acid residues including a putative signal peptide of 19 residues
and a mature toxin of 66 residues. They are different in 17 amino acid residues, among
them 11 residues in the mature toxin. The predicted amino acid sequences of BmK AS
and BmK AS-1 were almost consistent with those determined and revised (personal communication),
only different in one and two residues at their COO-terminal parts, respectively.
Based on the determined cDNA sequences, and using the total DNAs isolated from the
scorpion venom glands as a template, the genomic DNAs of BmK AS and BmK AS-1 were
also amplified by PCR and sequenced. It showed that no intron was inserted in their
open reading frames, while in the exon of signal peptide sequences of other Na+, K+
and Cl- channel toxins from the same scorpion, an intron is usually found. However,
the Northern blot hybridization results indicated that the sizes of their mRNA should
be around 800 bp. Their extra sequences around 400 bp which might function as an intron
should be located at their 5' untranslated regions.