Preparation of polyclonal antibody of N\|terminal peptide of cadherin of Helicoverpa armigera and primary detection of Btresistance.
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Key Words：Helicoverpa armigera, Ecadherin, prokaryotic expression, polyclonal antibody, Bt resistance
Abstract： Rabbit polyclonal antibodies were prepared with recombinant N\|terminal peptide of Helicoverpa armigera (Hübner) cadherin, and were used for identification of Bt\|resistance. N\|terminal peptide fragment encoding gene Cad285 was amplified by RT\|PCR from midgut of H. armigera and cloned to prokaryotic expression vector pET\|30a, then induced expressing in E. coli BL21 with IPTG. Recombinant protein (~ 35ku) was highly expressed and existed as inclusion bodies. The inclusion bodies were purified with denaturing, purifying by Ni\|NTA and refolding. The purified recombinant Cad285 was used to immunize rabbit for preparing polyclonal antibody with higher titer than 1∶16 000 measured by ELISA. Finally, a laboratory strains of H. armigera was detected by western blot with the polyclonal antibody. The results showed a significant difference between the sensitive and resistant strains.