苏云金杆菌制剂毒素蛋白定量分析_英文_刘丰茂.pdfVIP

1999, 1(2): 7381 Chinese Journal of Pesticide Science QUANTITATIVE ANALYSIS OF TOXIN PROTEINS IN BACILLUS THURIN GIENSI S PRODUCTS Liu Feng ao　Qian Chuanfan　Jiang Shuren (Depart ent of Applied Che istry, China Agricultural University, Key Laboratory of Pesticide Che istry and Application T echnology of M inistry of Agriculture, Beijing 100094) AbstractThe quantitative ethod, Sodiu Dodecyl Sulphate Polyacrya ide Gel Electrophoresis ( SDS- PAGE) -Colori etry, of toxin proteins in various B. t. co ercial products was presented. B . t. sa ples were pretreated with aqueous NaOH at high pH and treated with Lae, li sa ple buffer then electrophore-, sis was carried out. After stained with Coo assie Brilliant Blue (CBB) R-250, the band of 130KD-toxin pro- teins was cut out and the absorbed CBB R, -250 was eluted with pyridine The absorbance of the eluted dye. was easured at 605n . T his ethod characterized by relatively high accuracy and precision. Relative stan- dard deviation RSD was nearly( ) 5%. The fortified recoveries varied fro 94. 6%99. 2%. There were no significant differences between the datafro SDSColori etry and SDST LC-Scanner. T here- lationship between the percentage of 130KD-toxin protein and the potency of insect bioassay was studied. Key wordsBacillus thuringiensis;Quantitative analysis; Electrophoresis;Colori etry 1I troductio Bacillus thuringiensis (B.t.) is a species of gra positive aerobic soil bacteriu , which shows high toxicity to the larvae of various insects including Lepidoptera, Coleoptera and Diptera It is one of the bioinsecticide products which has been developed fastest and used. widest in the world. The first approach to assess thepotency of B.t. was to count the variablespores. Based on the ability of ost strains to produce one crystal and one spore, spore count could be re- lated to a ount of crystal and then