Abstract
The phosphinothricin acethl transferase(pat) gene was cloned and constructed into a prokaryotic expression vectorpET30(+)-pat.The protein expressed by this gene was purified and used as immunogen to immunize NewZealand rabbits and mice for polyclonal antibody and monoclonal antibody separately.The polyclonal antibody titer was greater than 8000,the monoclonal antibody(mAb 2F6) titer was 5 ×104.The mAb was IgG1 isotype with κ light chains.pat protein sandwich enzyme-linked immunosorbent assay was developed with anti-pat protein mouse monoclonal antibody(mAb) and rabbit polyclonal antibody.The linear range of the method was 1.56-100.0 μg/L.The linear equation was y=0.6914x-2.572,and the determinative coefficient was 0.9951.The assay was used to detect the content of pat protein in the leaf of five transgenic Bt cotton and two non-GM cotton varieties.The results showed that,three varieties of transgenic Bt cotton were positive and the others were negative.
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