BIOL207 Lecture Notes - Lecture 27: Lambda Phage, Sodium Hydroxide, Insulin
21 Apr 2018
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Joins 5" phosphate end to 3" h(cid:455)dro(cid:454)(cid:455)l end. Sticky end molecules are joined to form recombinant dna. Blunt end molecules do not ligate as efficiently as sticky end molecules. Sticky ends with non-complementary dna will not ligate. Repairing breaks in single or double stranded dna during recombination. Dna ligase facilitates the insertion of double stranded dna fragment into a plasmid vector. Essential to produce recombinant dna: basic terminology. In situ: experiments done on cells and structures removed from a live organism. In vitro: experiments done on molecules removed from an organism. In silico: experiments done within a computer simulation. Recombinant dna: dna molecule created in vitro by joining foreign dna with vector. Plasmids: small, circular pieces of dsdna that replicate independently of the chromosome. Often contain genes for pathogenicity and drug-resistance. Transformation: plasmids inserted into bacterial host cells. Both the foreign dna and plasmid are cut using restriction enzymes. Ligase joins compatible ends and form recombinant plasmid.
