| abstract |
The present invention is to produce a melitin antimicrobial peptide from silkworm body fluid using a silkworm transformation 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 for selection. Silkworm eggs were injected into 300 silkworm eggs to select 11 silkworm transformants in F1 generation. The selected silkworm transgenic plants were found to express EGFP fluorescent protein in the eyes and nervous tissues, larvae and pupae of adult stage, and adult eyes. In addition, G2 generation silkworm transformants were cultured 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 were crossbreeding each other. The G3 generation silkworm transformants selected in this way were tested for antimicrobial activity in the same manner as in the previous step. As a result, the antimicrobial activity was almost the same as that of Sigma's melittin (0.016 mg / ml) used as a control. Thus, it was confirmed from the above results that silkworm transformants producing melittin antimicrobial peptides were successfully produced. |