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In Molecular biology, an insert is a piece of DNA that is inserted into a larger DNA vector by a recombinant DNA technique, such as ligation or recombination. This allows it to be multiplied, selected, further manipulated or expressed in a host organism .
The transferred DNA is piloted to the plant cell nucleus and integrated into the host plants genomic DNA.The plasmid T-DNA is integrated semi-randomly into the genome of the host cell. [ 23 ] By modifying the plasmid to express the gene of interest, researchers can insert their chosen gene stably into the plants genome.
Recombinant DNA molecules are sometimes called chimeric DNA because they can be made of material from two different species like the mythical chimera. rDNA technology uses palindromic sequences and leads to the production of sticky and blunt ends. The DNA sequences used in the construction of recombinant DNA molecules can originate from any ...
These concepts were embodied in the model of DNA produced through the combined efforts of James Watson, Francis Crick, and Rosalind Franklin. Further research on the Watson-Crick model yielded theoretical advances that were reflected in new capacities to manipulate DNA. [6] One of these capacities was recombinant DNA technology.
Insert the fragments of DNA into vectors that were cut with the same restriction enzyme. Use the enzyme DNA ligase to seal the DNA fragments into the vector. This creates a large pool of recombinant molecules. These recombinant molecules are taken up by a host bacterium by transformation, creating a DNA library. [9] [10]
Molecular cloning takes advantage of the fact that the chemical structure of DNA is fundamentally the same in all living organisms. Therefore, if any segment of DNA from any organism is inserted into a DNA segment containing the molecular sequences required for DNA replication, and the resulting recombinant DNA is introduced into the organism from which the replication sequences were obtained ...
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 tissue are cut into small pieces and soaked in a fluid containing suspended Agrobacterium. The bacteria will attach to many of ...
If the delivered DNA construct contains a selectable marker, then stably transformed cells can be selected and cultured using tissue culture methods. For example, if the delivered DNA construct contains a gene that confers resistance to an antibiotic or herbicide, then stably transformed cells may be selected by including that antibiotic or ...