PJB-2006-83
IMMUNOASSAY-BASED APPROACH FOR DETECTION OF NOVEL BACILLUS THURINGIENSIS -ENDOTOXINS, ENTOMOCIDAL TO COTTON APHIDS (APHIS GOSSYPII) AND WHITEFLIES (BEMISIA TABACI)
KAUSAR MALIK AND SHEIKH RIAZUDDIN
Abstract
A new strategy, which is based on immunoassay, has been developed to screen locally isolated Bacillus thuringiensis (Bt). Delta endotoxins of three Bt strains (INS 2.13, HFZ 24.3 and GU 9.1) were prepared, electrophoresed on native PAGE and electroeluted the purified delta endotoxins from gels. Determined the toxicity of purified delta endotoxins against cotton aphids (Aphis gossypii) and whiteflies (B. tabaci). N-terminal sequences of the purified proteins (d endotoxins) of these three entomocidal Bt isolates showed no significant homology to protein sequences in the Gen bank (NCBI) protein database. The purified proteins having novelty in their sequences were used as antigens for the immunization of rabbits to raise polyclonal antibodies. The affinity purified antibodies were used in immunoassays to screen 170 Bt isolates for entomocidal activity and obtained promising results.
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