| In this paper, The spore /crystal complex was prepared from Bacillus thuringiensis var. kurstaki HD-1, HD-73 and genetically engineered strain 304A(b) in 171 K EM+ during sporulation as crystalline inclusions that are releases along with spores. The parasporal crystal was isolated from the spore crystal/complex by liquid two phase method. CrylA, CrylA(b) and CrylA(c) were precipitated from parasporal crystal on PI4.4..3 polyclonal antibodies were acquired using CrylA, CrylA(c) and CrylA(b) as antigens. The double diffusion titer of anti-CrylA, Anti-CrylA(c) and CrylA(b) were 1/32, 1/16, 1/16 separately. ELISA titer of the 3 antisera were＞1/100000,＞1/100000,＞1/100000. 11 monoclonal antibodies were produced using CrylA insecticidal Crystal protein to immune BALA/C mouse. After testing the specificity of monoclonal antibody and polyclonal antibody by samples, only one clone B4E6D8, Anti- CrylA and Anti-CrylA(c) antibodies could be used.Three kinds of Bt toxin ELISA KITS were established. The detection sensitivity was less than 10 ng/ml. We detect a large numbers of transgenic Bt cotton samples, the negative samples were found correctly in every test. The result was identical with the field insect-resistant performance.It is very impportant to develop ELISA kits to detect and quantitate the levels of toxin expressed in transgenic Bt cotton. The assays can be used in insect management, seed quantity control and insect-resistant hybrid breeding.
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