Answer to Question #109201 in Molecular Biology for kylie

Question #109201
Recombinant DNA technology has had a dramatic impact on all aspects of molecular biology, allowing scientist to routinely study cells and their macromolecules in ways that were unimaginable. Describe manipulations central to this technology
1
Expert's answer
2020-04-20T09:49:44-0400


Recombinant DNA technology, also molecular cloning, or genetic engineering

include the following steps:


1. Restriction digestion from the donor organism of the necessary genes of native DNA (cloned DNA, inserted DNA, target DNA, foreign DNA).


2. Quick decoding of all nucleotides in the purified DNA fragment, which allows to determine the exact boundaries of the gene and the amino acid sequence encoded by the gene.


3. Treatment with restriction endonucleases of a cloning vector that can replicate in the host cell.


4. Crosslinking of two DNA fragments by DNA ligase with the formation of a new recombinant molecule - the “cloning vector - integrated DNA” construct.

5. The introduction of this construct into the host cell (recipient), where it is replicated and transmitted to descendants. This process is called transformation. After transformation, the bacterial cell reproduces a fragment of the cloned DNA by millions of identical cells.


6. Identification and selection of cells carrying recombinant DNA (transformed cells).


7. Obtaining a specific protein product synthesized by host cells


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