Buthus Scorpion Part Of The Toxin-genome Multiple Sequence Alignment Analysis

The genomic sequences of the Buthus martensii Karsch toxins was collected and saved as Fasta format. Sequences were appended to the ClustalX1.83 software, did complement alignment, saved as '*.aln' file. Converted the '*.aln' file to MEGA format, and opened data, exported data, constructed phylogeny and did computer. The result shows that the sodium channel toxins on the one hand and the potassium channel toxins on the other hand are two distinct principle branches of the unrooted phylogenetic tree. Calcium channel toxins lies in the edge of the potassium channel toxins. Cl~- channel toxins are hiding in the potassium channel toxins' branch.Genomic DNA was purified form the muscle tissue of the Buthus martensii Karsch scorpion. Cells from the muscle tissue were lysed by proteinase K and SDS, and the sample was gently extracted with phenol. DNA was precipitated by adding ammonium acetate and ethanol. Agarose gel electrophyrisis showed that the DNA's size was about 20kb. The genomic DNA of AGAP2 mature peptide was amplified under step down PCR reaction. PCR products were ligated into the pET28 vector. E.coli DH5αwas used for plasmid propagation. Recombinant clones were analyzed by PCR following agarose gel electrophoresis and SSCP. Compared with the amino acid sequence and the cDNA sequence of AGAP2, there is no intron in the mature peptide coding region.