[Site-directed mutagenesis of melittin gene and its expression in Escherichia coli]

The cDNA encoding promelittin was obtained from the total RNA of bee poison gland by RT-PCR. Moreover, hydroxylamine clearage site was arranged before the melittin sequences by site-directed mutagenesis. The expression vector containing the mutagenic promelittin sequence with partial sequence of beta-galactosidase was constructed. The result of DNA sequence analysis demonstrated that the obtained cDNA sequence include the desired codon and the reading frame of fusion gene was correct. The induced protein was expressed in Escherichia coli.