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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.
National Center of Excellence in Molecular Biology, University of the
Punjab, 87-West Canal Bank Road, Thokar Niaz Baig, Lahore-53700, Pakistan.
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