51 results about "Ti plasmid" patented technology

This invention relates to several plasmids which are useful for genetically transforming plant cells. A first plasmid, such as pMON120, contains a T-DNA border, one or more marker genes, a unique cleavage site, and a region of Ti plasmid homology. A gene which is expressed in plant cells may be inserted into this plasmid to obtain a derivative plasmid, such as pMON128 which expresses neomycin phosphotransferase in plant cells. The derivative plasmid is inserted into a suitable microorganism, such as A. tumefaciens which contains a Ti plasmid. The inserted plasmids recombine with Ti plasmids to form co-integrate plasmids. Only a single crossover event is required to create the desired co-integrate plasmid. A. tumefaciens cells with co-integrate plasmids are selected and co-cultured with plant cells. The co-integrate Ti plasmids enter the plant cells and insert a segment of T-DNA which does not contain tumorigenic genes into the plant genome. The transformed plant cell(s) may be regenerated into a morphologically normal plant which will pass the inserted gene(s) to its descendants.

The invention relates to a dual-elemental Ti plasmid carrier for the transformation of T-DNA (Transferred-Deoxyribose Nucleic Acid) of plants and partial fungi and the construction of a T-DNA insertion mutant library and a construction method of the dual-elemental Ti plasmid carrier. The method comprises the following steps: firstly, performing module trapping on a promoter integrated with sGFP (Green Fluorescent Protein) reporter genes at two ends of a T-DNA area of a plasmid; optimizing and adjusting a nucleotide sequence before opening reading frames for the sGFP at two ends of the nucleotide sequence; secondly, integrating a functional module rescued by the plasmid into a target plasmid through an enzyme cutting site. Finally, the plasmid is integrated with a hygromycin resistance gene which is used for screening a transformant; the hygromycin resistance gene and the promoter thereof can be replaced freely through a Sac I enzyme cutting site so as to adapt to different screening purposes and species. The convenience in the efficiency, the gene cloning and the related gene function verification of transgene is seriously affected by the selection and the module configuration and optimization of the dual-elemental Ti plasmid carrier. The application range of the plasmid is enlarged and the screening and cloning efficiency of target functional genes is improved by organically integrating various plasmid functional modules into one plasmid. Thus, a good molecular tool for finding beneficial genes and verifying the functions of the beneficial genes is provided.