give and illustrated in account of gene cloning
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Gene Cloning
Gene cloning is a commonly used molecular biological technique in which a gene of interest is fused into a self-replicating genetic element called a plasmid, which when introduced into a suitable host (usually bacteria), self-replicates and generates a large number of identical copies of the particular gene.
In a regular molecular cloning experiment, the DNA containing the desired gene is isolated from the organism. This DNA is cut into the right size using restriction enzymes. The plasmid (which is used as the vector) is also treated using the same restriction enzymes. Cutting with the same restriction enzymes ensures that the ends of the vector fragment are compatible with the ends of the DNA fragment that needs to be cloned. Next, the DNA fragment and the vector are mixed together and allowed to ligate together. Ligation is performed using ligase enzymes. The recombinant plasmid, which now contains the DNA fragment as an insert, is transferred into the bacteria through a process called transformation. The bacteria are then grown on growth media containing specific antibiotics that allow only plasmid containing bacteria to grow (selection). The bacteria are grown in large numbers, and plasmids are isolated from them. Each of these plasmids would carry the gene of interest as the insert, which if needed can be isolated using the same restriction enzyme that was initially used to insert them into the vector.
Gene cloning is a commonly used molecular biological technique in which a gene of interest is fused into a self-replicating genetic element called a plasmid, which when introduced into a suitable host (usually bacteria), self-replicates and generates a large number of identical copies of the particular gene.
In a regular molecular cloning experiment, the DNA containing the desired gene is isolated from the organism. This DNA is cut into the right size using restriction enzymes. The plasmid (which is used as the vector) is also treated using the same restriction enzymes. Cutting with the same restriction enzymes ensures that the ends of the vector fragment are compatible with the ends of the DNA fragment that needs to be cloned. Next, the DNA fragment and the vector are mixed together and allowed to ligate together. Ligation is performed using ligase enzymes. The recombinant plasmid, which now contains the DNA fragment as an insert, is transferred into the bacteria through a process called transformation. The bacteria are then grown on growth media containing specific antibiotics that allow only plasmid containing bacteria to grow (selection). The bacteria are grown in large numbers, and plasmids are isolated from them. Each of these plasmids would carry the gene of interest as the insert, which if needed can be isolated using the same restriction enzyme that was initially used to insert them into the vector.
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Gene cloning is the process in which a gene of interest is located and copied (cloned) out of DNA exctrated from an organism .When DNA is extracted from a organisms,all of its genes are exctrated at one time.This DNA which contains thousands of difference genes.
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