The gene cloning and sequencing of Bm-12, a chlorotoxin-like peptide from the scorpion Buthus martensi Karsch.
According to the known amino acid sequence of Bm-12, a short chain insect neurotoxin from the venom of the scorpion Buthus martensi Karsch (BmK) with considerable primary sequence homology to chlorotoxin, the gene specific primers were designed and synthesized for 3' and 5'RACE (Rapid Amplification of cDNA Ends). The two partial cDNA fragments obtained by 3' and 5'RACE were cloned and sequenced, and the full length cDNA sequence of Bm-12 was then completed by overlapping these two partial cDNA sequences. The predicted amino acid sequence consists of 59 amino acid residues including a putative signal peptide of 24 residues and a mature toxin of 35 residues. The predicted amino acid sequence of Bm-12 was almost consistent with the determined, different only in one residue at position 27, Lys was replaced by Gly. Based on the determined cDNA sequence, and using the total DNA isolated from the scorpion venom glands as a template, the genomic DNA of Bm-12 was also amplified by PCR and sequenced. The genomic DNA sequence revealed an intron of 93 bp present within the signal peptide region.
Wu
J J
JJ
State Key Laboratory of Molecular Biology, Shanghai Institute of Biochemistry, Academia Sinica, People's Republic of China.
Dai
L
L
Lan
Z D
ZD
Chi
C W
CW
D000595:Amino Acid Sequence; D000818:Animals; D001097:Arachnida; D001483:Base Sequence; D003001:Cloning, Molecular; D004247:DNA / Q000032:analysis*; D017931:DNA Primers / Q000737:chemistry; D005784:Gene Amplification; D008969:Molecular Sequence Data; D009498:Neurotoxins / Q000737:chemistry / Q000235:genetics*; D012313:RNA / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012604:Scorpion Venoms / Q000737:chemistry / Q000235:genetics*; D012605:Scorpions / Q000235:genetics*; D017386:Sequence Homology, Amino Acid
D017931:DNA Primers; D009498:Neurotoxins; D012604:Scorpion Venoms; C091539:Chlorotoxin; D012313:RNA; D004247:DNA