10 - Kuo, 580.221 10 Oct 2006 Concepts/Vocabulary Molecules...

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Kuo, 580.221 10 Oct 2006 1 BME 580.221 Generation of Antibody Diversity Lecture #10, 10 Oct 2006 Scot C. Kuo 10 Oct 2006 Kuo, 580.221 Concepts/Vocabulary • Clonal Selection Hypothesis -- ‘cellular’ RARE somatic (DNA) rearrangement: VDJ recombination – Contrast with splicing (RNA), genomic recombination, biotechnology (“splice”, “recombinant”) • Southern blot (and Northern/Western) 10 Oct 2006 Kuo, 580.221 Revisit Cowpox vaccination Two immunological roles of viruses : Immunogen (stimulate immune response, cowpox) vs. Antigen (target of response, smallpox) In variolation, smallpox virus serves as both immunogen and antigen. The first half of the course will explain the chemical, genetic, and cellular mechanisms underlying the clonal selection hypothesis . Clonal Selection Hypothesis (see readings; more in later lectures) 10 Oct 2006 Kuo, 580.221 Clonal Selection Hypothesis: “memory” at cellular level Key features (later list more detailed) • Each Ab-producing cell produces only one Ab specificity – specificity pre-exists encountering Ag • Ab “displayed” on surface of cells – binding of Ag stimulates cell growth (proliferation) and Ab secretion • Clones persist after disappearance of antigen (memory) 10 Oct 2006 Kuo, 580.221 Problem with accepting Clonal Selection Hypothesis -- too many antibodies • First -- review Dogma of Genetic Information in Molecular Biology (modified with extra processes) • Reminder: Dogma has been extremely successful – very reasonable framework to start. 10 Oct 2006 Kuo, 580.221 Central Dogma of Molecular Biology • DNA->RNA->protein Bacterial paradigm extremely successful. Conventional wisdom : • DNA is reference (sancrosanct) • one gene = one protein
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Kuo, 580.221 10 Oct 2006 2 10 Oct 2006 Kuo, 580.221 Protein Expression in Eukaryotes More complex than bacteria, but we can still have ways to rationalize the extra reactions: Why have ‘useless’ information, i.e. introns ? Why have a nucleus? Why have polyadenylation? NOTE: All are POOR questions for the scientific method! 10 Oct 2006 Kuo, 580.221 RNA maturation: ‘primary’ transcript -> mRNA (pre-mRNA=hnRNA) 10 Oct 2006 Kuo, 580.221 Proof Splicing Exists: RNA-DNA hybridization of Adenovirus 10 Oct 2006 Kuo, 580.221 Splicing is ONLY with RNA (why?) Why is DNA for genome (permanent) and RNA for working copies? 10 Oct 2006 Kuo, 580.221 Splicing (view from pre-mRNA) 10 Oct 2006 Kuo, 580.221 Processes in Molecular Biology, complex (Open reading frame) Glycosylation Splicing, PolyAdenyl., Enhancers, RevTranscriptase Telomeres, Methylation, Histones Eukaryotic, Special Stop Start Enzymes Subunit Process, Directionality Polym. Stop
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10 - Kuo, 580.221 10 Oct 2006 Concepts/Vocabulary Molecules...

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