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 ultimate information regulation Pharmaceutical manufacture medicines Clinical

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!!! 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 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 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: amp R gene and lacZ+ gene

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)