[Cloning of promelittin cDNA and its expression in Escherichia coli]

The cDNA encoding promelittin was obtained from the total RNA of bee poison gland by RT-PCR and was cloned to pT7Blu-T vector. The expression vector of promelittin fused with partial sequence of beta-galactosidase was constructed by ligating the fragment inserted to pUC118. Moreover, it was expressed in the strain DH5 alpha of Escherichia coli. The result of DNA sequence analysis demonstrated that the obtained cDNA sequence was same with the published one and the reading frame of fusion gene was correct.