FIELD: chemistry; biochemistry.
SUBSTANCE: plant explants are co-cultured with Agrobacterium tumefaciens bacteria strain which contains recombinant plasmid pBM which bears the gene of the desired product and free from selective markers of antibotic and herbicide resistance genes. The transformed explants are put into a medium which does not contain antibiotics to initiate sprouting. The obtained sprouts are grown in fresh medium to fertile transgenic plants which bear the target gene and having features of the initial sort, and the plants are directly selected based on the desired products they have synthesised. The strain Agrobacterium tumefaciens LBA4404 (pAL4404) is used for transformation. Transformants are selected after regeneration of sprouts through immunoenzymometric or western-blot analysis directly, analysing extracts of seedlings formed in the culture medium which does not contain selective antibiotics and which express the gene of the desired product. The gene of the desired product is selected from a group comprising a gene which codes the hepatitis B virus antigen, a gene which codes the antimicrobial peptide of mammal cecropin P1, the gene of delta endotoxin Bacillus thuringiensis CrylA(b) and the gene of bacterial DNA methyltransferase M.EcoRIL.
EFFECT: method enables to obtain non-marker transgenic plants.
7 cl, 3 dwg, 17 ex
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Authors
Dates
2011-01-27—Published
2009-06-02—Filed