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Q.Give a brief account of the tools of recombinant DNA technology.

Telangana TsbieTelangana Board of Intermediate Education 2023Subjective· 8mImportance★★★★★
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The key tools of rDNA technology are restriction enzymes (cut), DNA ligase (join), cloning vectors (carry and replicate), and host/competent cells (multiply the DNA), aided by polymerases (PCR) and gel electrophoresis.

Genetic engineering (recombinant DNA technology) requires certain essential tools:

  1. Restriction enzymes: These are 'molecular scissors'. Restriction endonucleases recognise specific palindromic sequences in DNA and cut both strands, often leaving single-stranded overhangs called sticky ends. Example: EcoRI. The sticky ends allow the DNA of different sources to be joined. (Exonucleases remove nucleotides from the ends; endonucleases cut within.)

  2. DNA ligase: This is the 'molecular glue' that joins the DNA fragments (e.g. the vector and the foreign DNA with complementary sticky ends) by sealing the sugar-phosphate backbone.

  3. Cloning vectors: These are DNA molecules (such as plasmids and bacteriophages) that carry the foreign DNA into the host and replicate within it. A good vector must have: origin of replication (ori), a selectable marker (e.g. antibiotic resistance genes to identify transformants), and unique recognition sites for restriction enzymes. Examples: the plasmid pBR322, and the Ti plasmid of Agrobacterium for plants.

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