[Screening and rapid identification of Bacillus thuringiensis mutants].
Mutants of Bacillus thuringiensis subsp. kurstaki NTU 9 and Bt 158, which were isolated previously for using the diamondback moth as a target insect in Taiwan, were screening by either protein electrophoresis of intracellular proteins or enzyme-linked immunosorbent assay (ELISA). The optimal conditions of effective protein electrophoresis were (1) 24-hour cells harvested from nutrient broth were crashed by petite glass beads followed by centrifugation. And (2) the supernatant pretreated by heating at 60 degrees C for 2 minutes was electrophoresed with 7.5% native PAGE at 110 voltages. On ELISA, the antiserum used was obtained from rabbits immunized with Bt 158 crystal protein. Optimal antigen coating concentration of ELISA, attained by chequer-board titration method, was 10 micrograms/ml. Antigens (crystal protein) in samples were detected by competitive inhibition method with antiserum diluted to 10(4) fold. By using protein electrophoresis and ELISA methods, two isolates A 71 and BN 11, were denoted respectively as qualitative and quantitative mutants of Bacillus thuringiensis.
Su
Y C
YC
Graduate Institute of Agricultural Chemistry, National Taiwan-University, Taipei, R.O.C.
Lee
S F
SF
Chiou
S Y
SY
D000818:Animals; D000942:Antigens, Bacterial / Q000032:analysis; D001413:Bacillus thuringiensis / Q000737:chemistry / Q000235:genetics* / Q000276:immunology; D001426:Bacterial Proteins / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D009154:Mutation; D011817:Rabbits
D000942:Antigens, Bacterial; D001426:Bacterial Proteins