Preparation of polyclonal antibodies against acetolactate synthase of Lolium multiflorum and its application in detection of various weeds
In order to achieve serological detection of the target enzyme ALS of weeds and enrich the detection methods of this enzyme,the ALS gene of Lolium multiflorum was ligated into the prokaryotic expression vector pGEX-2TK,the recombinant expression plasmid pGEX-2TK-Lm ALS was transformed into Escherichia coli BL21 cells,and the recombinant fusion protein GST-LmALS was induced and purified.After immunizing mice with purified fusion protein as immunogen for four weeks,anti-ALS polyclonal antibody was obtained.The titer of the antibody was determined by indirect ELISA,and the results showed that the antibody had a good titer,which could reach 1∶256 000.Western blot analysis demonstrated that the prepared anti-ALS polyclonal antibody can specifically detect the ALS of L.multiflorum,and can also detect the ALS of various weeds.Preparation of polyclonal antibodies will lay a solid foundation for the subsequent study on the biochemical characteristics of herbicidal target enzyme ALS.
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