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Successful gene delivery requires the foreign gene delivery to remain stable within the host cell and can either integrate into the genome or replicate independently of it. [3] This requires foreign DNA to be synthesized as part of a vector , which is designed to enter the desired host cell and deliver the transgene to that cell's genome. [ 4 ]
There are several binary vectors that replicate in Agrobacterium and can be used for delivery of T-DNA from Agrobacterium into plant cells. The T-DNA portion of the binary vector is flanked by left and right border sequences and may include a transgene as well as a plant selectable marker.
The ability of Agrobacterium to transfer genes to plants and fungi is used in biotechnology, in particular, genetic engineering for plant improvement. Genomes of plants and fungi can be engineered by use of Agrobacterium for the delivery of sequences hosted in T-DNA binary vectors. A modified Ti or Ri plasmid can be used.
To interact with host plant proteins many Agrobacterium virulence proteins encoded by vir genes. Agrobacterium vir gene expression occurs via the VirA-VirG sensor that results in generation of a mobile single-stranded T-DNA copy (T-strand). A processed form of VirB2 is the major component of the T-complex that is required for transformation.
A number of methods are available to transfer DNA into plant cells. Some vector-mediated methods are: Agrobacterium-mediated transformation is the easiest and most simple plant transformation. Plant tissue (often leaves) are cut into small pieces, e.g. 10x10mm, and soaked for ten minutes in a fluid containing suspended Agrobacterium. The ...
A gene gun uses biolistics to insert DNA into plant tissue. A. tumefaciens attaching itself to a carrot cell. In plants the DNA is often inserted using Agrobacterium-mediated recombination, [27] taking advantage of the Agrobacteriums T-DNA sequence that allows natural insertion of genetic material into plant cells. [28]
Plant transformation vectors are plasmids that have been specifically designed to facilitate the generation of transgenic plants.The most commonly used plant transformation vectors are T-DNA binary vectors and are often replicated in both E. coli, a common lab bacterium, and Agrobacterium tumefaciens, a plant-virulent bacterium used to insert the recombinant DNA into plants.
Genetically modified crops (GM crops) are plants used in agriculture, the DNA of which has been modified using genetic engineering methods. Plant genomes can be engineered by physical methods or by use of Agrobacterium for the delivery of sequences hosted in T-DNA binary vectors.