Dna cloning
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DNA Cloning. Cloning. a line of genetically identical cells or individuals derived from a single ancestor produces many copies of a piece of DNA uses a little fraction as gene of interest cultivates a large amount for studying functions. Functions. Biological determine the DNA sequence

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DNA Cloning

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Dna cloning

DNA Cloning


Cloning

Cloning

  • a line of genetically identical cells or individuals derived from a single ancestor

  • produces many copies of a piece of DNA

  • uses a little fraction as gene of interest

  • cultivates a large amount for studying functions


Functions

Functions

  • Biological

  • determine the DNA sequence

  • ultimate information

  • regulation

  • Pharmaceutical

  • manufacture medicines

  • Clinical


How it works

How it Works

  • Combine gene of interest and bacterial plasmid

  • Recombinant DNA

  • DNA from two or more different sources that have been joined together to form a single molecule

  • Amplification and identification


Zoom in

Zoom In!!!

  • Insertion, the most critical step

  • made possible by restriction endonucleases(aka. restriction enzymes)

  • recognize specific DNA sequences called restriction sites

  • cut the DNA at specific locations

  • Restriction fragments


Restriction enzyme

Restriction Enzyme

  • defend against viral attack by breaking down the DNA molecules of infecting viruses

  • cleave the sugar-phosphate backbones of DNA to produce sticky ends

  • short single-stranded regions

  • form hydrogen bonds with complementary sticky ends on any other DNA molecules cut with the same enzyme

  • kind of like glue when you fix something


Cloning vectors

Cloning Vectors

  • DNA molecules into which a DNA fragment can be inserted to form a recombinant DNA molecule for the purpose of cloning

  • plasmid cloning vectors

  • contains 2 unique genes: ampR gene and lacZ+ gene


The two genes

The Two Genes

  • ampR gene

  • encodes an enzyme that breaks down the antibiotic ampicillin

  • lacZ+ gene

  • encodes ß-galactosidase (hydrolyzes the sugar lactorse and synthetic substrates)


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