Study On The Separation Of Mythimna Separata Midgut Binding Protein With The Method Of Dimethyl Anthranilat Labeling Periplocoside P

In recent years, the active ingredient and its insecticidal effect of Periploca sepium Bunge have carried in detail by Institution of Pesticide, Northwest A&F University. By the different treatments of protein with drugs which are in vitro incubation and in vivo feeding, we separated and purified the binding protein of periplocoside P from the midgut of Mythimna separata larvae, and it’s a preliminary exploration work to study the target protein. The main results are as follows:(1) Modified the periplocoside P with the N-Methylisatoic anhydride, and got the labeling production of periplocoside P-MIA. Bioactivity assay to test the insecticidal activities of the periplocoside P-MIA were reported the poisoning symptoms are the same as periplocoside P. By feeding with 5μg drugs on the 0.5 cm x 0.5 cm wheat leaf disc, 48 h mortality was 12.5%, lower than the activity of periplocoside P in the same condition.(2) To analysis two different treatments of drugs and crude protein in vitro incubation and in vivo feeding with Q-Sepharose anion exchange column chromatography, we monitored four ultraviolet absorption chromatograms of different protein treatment groups(control group, periplocoside P group, periplocoside P-MIA group, dimethyl anthranilat group) under the UV wavelength of 350 nm. The different absorption value protein fractions were used to SDS-PAGE. By the protein mass spectrometry analysis, we get 8 kinds of binding proteins of periplocoside P, there are abnormal wing disc-like protein, nucleoside diphosphate kinase, alcohol dehydrogenase, partial, methylmalonate-semialdehyde dehydrogenase, slingshot dual specificity phosphatas, 2OG-Fe(2)oxygenase superfamily protein, fructose1, 6-bis-phosphate aldolase, Catalase.(3) To analysis two different treatments of drugs and BBMV in vitro incubation and in vivo feeding with Q-Sepharose anion exchange column chromatography, we monitored four ultraviolet absorption chromatograms of different protein treatment groups(control group, periplocoside P group, periplocoside P-MIA group, dimethyl anthranilat group) under the UV wavelength of 350 nm. The different absorption value protein fractions were used to SDS-PAGE. By the protein mass spectrometry analysis, we get 10 kinds of binding proteins, there are abnormal wing disc-like protein, 2OG-Fe(2)oxygenase superfamily protein, aminopeptidase N, catalase, double-stranded RNA-binding protein Zn72 d, NADH-ubiquinon-eo xidoreductase 75 kDa subunit, protein disulfide isomerase, arylphorin and Arginine kinase-like protein.