LIFESCI 3 Lecture Notes - Lecture 19: Multiple Cloning Site, Genomic Library, Dna Ligase
Document Summary
Background: recombinant dna, definition: any synthesized dna molecule that wouldn"t normally exist in nature, most common purpose: gene cloning, coli immune system . Modification enzymes: methylase, strain specific, protects strain"s genome from restriction endonuclease cleavage by methylating a specific sequence. Essential components of vectors: definition: dna that is introduced into the vector, selectable marker for bacteria containing the plasmid, ex. Other components of vectors: polylinker, also called a multiple cloning site (mcs, definition: synthesized dna sequence that usually contains 10-20. Shine dalgarno sequence: the plasmids are sometimes inducible, depending on the regulatory region that"s controlling the rate of transcription, can turn it on only in the presence of certain molecules or factors, ex. Process: (1) on foreign gene that will be cloned, check sequences on both sides of it for what restriction enzymes it contains, the restriction enzymes have to be pretty close to flanking the 5".
