Production of the melittin antimicrobial peptide in transgenic silkworm

摘要

The present invention is to produce a melittin antimicrobial peptide from silkworm body fluid using a silkworm transfection technique. In this experiment, the melittin antimicrobial peptide was expressed using the actin-3 promoter derived from silkworm. For selection of silkworm transformants, 3xP3 promoter and EGFP gene were used to select silkworm eggs, and 11 silkworm transformants were selected from F1 generations by injection into 300 silkworm eggs. The selected silkworm transformants were found to express the EGFP fluorescent protein in the eyes and nervous tissues of the early embryonic stage, larvae and pupae, and adult eyes. G2 generation silkworm transgenic plants were raised to 5th instar 5th day larvae, and body fluids were collected and pretreated. The samples were tested for antimicrobial activity, and a total of 10 silkworms were selected. These selected silkworms crossed each other by crossing. The G3 generation silkworm transformants selected in this way were tested for antimicrobial activity in the same manner as described above. As a result, the antimicrobial activity was almost the same as that of Sigma's melittin (0.016 mg / ml) used as a control. As a result, it was confirmed that silkworm transformants producing melittin antimicrobial peptides were successfully produced.;