MMG301 1nd Edition Lecture 25 Outline of Last Lecture I. Bacterial geneticsOutline of Current Lecture II. Biotechnology III. PCRIV. plasmidsCurrent Lecture-joining DNA from different origins yields recombinant DNA-a large population of heterogeneous fragments (called a library) can be joined into a single typeof cloning vector.-restriction enzymes (restriction endonucleases)-recognize a 4-8 base pair (bp) sequence in dsDNA-make a cut in both strands of the DNA-fragments are joined using an enzyme call DNA ligase-uses for PCR: -sequencing/evolutionary relationships – 16S RNA sequencing-diagnostics – detection of microbial infections-detection of viable but non-culturable microbes-plasmids: pUC19 – used for many years-ampicillin resistance (b-lactamase) used to select only for cells that contain plasmid-origin of replication-many restriction enzyme site for cloning DNA fragments• Other cloning plasmids are used for specific purposes:These notes represent a detailed interpretation of the professor’s lecture. GradeBuddy is best used as a supplement to your own notes, not as a substitute.-artificial chromosomes -yeast (YAC): propagate like a separate chromosome; large DNA inserts-bacterial (BAC): based on E. coli F plasmid; inserts up to 300 kbp-expression plasmids: Designed with restriction enzyme sites and transcription and translation elements to maximize production of recombinant proteins-shuttle vector - has origin of replication for eukaryotic organism (yeast, insect cells) AND an origin of replication for E.
View Full Document
Unlocking...