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Plasmid miniprep. 0.8% agarose gel ethidium bromide-stained.. A plasmid preparation is a method of DNA extraction and purification for plasmid DNA.It is an important step in many molecular biology experiments and is essential for the successful use of plasmids in research and biotechnology.
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.
These plasmid are generally non-conjugative but may have many more features, notably a "multiple cloning site" where multiple restriction enzyme cleavage sites allow for the insertion of a transgene insert. The bacteria containing the plasmids can generate millions of copies of the vector within the bacteria in hours, and the amplified vectors ...
This process can be done in several ways, depending on the type of the sample and the downstream application, [3] the most common methods are: mechanical, chemical and enzymatic lysis, precipitation, purification, and concentration. The specific method used to extract the DNA, such as phenol-chloroform extraction, alcohol precipitation, or ...
The pGreen plasmids are vectors for plant transformation. They were first described in 2000 as components of a novel T-DNA binary system. [1] The supporting web page [2] provides supplementary information and ongoing support to researchers to request their plasmid resources. As these plasmids have been taken up by the research community, the ...
The vir helper plasmid contains the vir genes that originated from the Ti plasmid of Agrobacterium. These genes code for a series of proteins that cut the binary vector at the left and right border sequences, and facilitate transfer and integration of T-DNA to the plant's cells and genomes, respectively. [4]
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. [29] By modifying the plasmid to express the gene of interest, researchers can insert their chosen gene stably into the plants genome.
The term plasmid was coined in 1952 by the American molecular biologist Joshua Lederberg to refer to "any extrachromosomal hereditary determinant." [14] [15] The term's early usage included any bacterial genetic material that exists extrachromosomally for at least part of its replication cycle, but because that description includes bacterial viruses, the notion of plasmid was refined over time ...
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