“Bacterial” genetics ISlide 2DNA RNA proteinSlide 4Slide 5Slide 6Slide 7Let’s all do science in NevadaSlide 9Slide 10What Luria actually didS. Luria, M. Delbrück (1943) Mutations of bacteria from virus sensitivity to virus resistance. Genetics 28: 491-511.Slide 13Slide 14Slide 15Slide 16Slide 17Joshua Lederberg“Conjugation” in bacteriaJ. Lederberg, E. Tatum (1946) Novel genotypes in mixed cultures of biochemical mutants of bacteria. Cold Spring Harbor Symp. Quant. Biol. 11: 113-114.Slide 21B.H. and A.H.William HayesSelman WaksmanSlide 25Rich husband, poor wife is not the same as poor husband, rich wifeSlide 27Slide 28Slide 29Slide 30Slide 31Slide 32Slide 33Slide 34Slide 35Cours de Physiologie et de génétique bactériennes, 1957 Illustration humoristique des cours par une série de dessins de F. Lavallé Légendes de Georges Cohen http://www.pasteur.fr/infosci/archives/mon/im_laval.htmlThe life cycle of a temperate phageThree aspects of phage biology with long-term impactSlide 39Slide 40Slide 41Slide 42Slide 43Slide 44Slide 45Slide 46Further readingMCB 140 2/16/05 1“Bacterial” genetics IMCB 140 2/16/05 2“What are the genes? What is the nature of the elements of heredity that Mendel postulated as purely theoretical units? … Frankly, these are questions with which the working geneticist has not much concern himself… If the gene is a material unit, it is a piece of a chromosome; if it is a fictitious unit, it must be referred to a definite location in a chromosome. … Therefore, it makes no difference in the actual work in genetics which point of view is taken.”T.H. Morgan The Relation of Genetics to Physiology and MedicineNobel Lecture, June 4, 19343MCB 140, 2/25/05DNA RNA proteincentral dogma of molecular biologyMCB 140 2/16/05 47.17MCB 140 2/16/05 5Max Delbrück (1906 -1981) & Salvador Luria (1912 - 1991), Cold Spring Harbor Laboratory, Long Island NY, Summer 1941MCB 140 2/16/05 6Today even the layman thinks of resistant bacteria as originating from mutation … but when Luria and Delbrück first got together, conventional bacteriologists were by no means clear that microorganisms could be tought about genetically… Many believed that resistance was some kind of adaptation induced, in a few of the bacteria in a culture, by the exposure to the antibacterial agent.Judson p. 55MCB 140 2/16/05 7The idea smacks of the pre-Mendelian, pre-Darwinian notion of the inheritance of acquired characteristics; Luria damned bacteriology as the “last stronghold of Lamarckism.”Judson p. 55MCB 140 2/16/05 8Let’s all do science in NevadaOne Saturday evening … Luria went to a faculty dance… There, watching the fluctuating returns obtained by colleagues gambling on a slot machine, he thought of the experiment that would distinguish between resistance induced in bacteria and resistance resulting from previous spontaneous mutation upon which selection acts. Judson p. 55MCB 140 2/16/05 9What Luria perceived was that previous spontaneous mutation would pay out jackpots of resistant bacteria that would fluctuate much more widely in size than those paid out by induction. He tried the first experiment on the following morning and wrote off to Delbrueck; Delbrueck promptly replied that Luria really ought to go to church …Judson p. 55MCB 140 2/16/05 107.4MCB 140 2/16/05 11What Luria actually didSample set A:1. Inoculate bacteria into individual cultures (1 bacterium per culture).2. Let it grow up to a large number.Sample set B:1. Take an aliquot of bacteria, and start a culture (which will therefore not be clonal).2. Let them grow up to a large numberExpose both to phage, and count, how many phage-resistant colonies per culture are found. Ask, if there is a difference between these two sample sets.MCB 140 2/16/05 12S. Luria, M. Delbrück (1943)Mutations of bacteria from virus sensitivity to virus resistance. Genetics 28: 491-511.“If the production of resistance began only at the moment of exposure to phage, then it wouldn’t matter whether the bacteria came from many individual cultures or one bulk culture. … When Luria performed the experiment, though, the twenty separate cultures showed much wider fluctuations from the average number of resistant colonies, indicating that a few of the individual tubes contained resistant bacteria from near the beginning of the overnight growth period.”Judson p. 56MCB 140 2/16/05 13Brock p. 59MCB 140 2/16/05 14MCB 140 2/16/05 15MCB 140 2/16/05 16George Beadle (left) and Edward Tatum (right) receiving their Nobel PrizesMCB 140 2/16/05 17MCB 140 2/16/05 18Joshua LederbergMCB 140 2/16/05 19“Conjugation” in bacteriaTake strain of E. coli that is auxotrophic for two distinct nutrients (thiamine and leucine).Take different strain of E. coli that is also auxotrophic for two distinct nutrients, but different ones (biotine and cysteine).Mix the two.Ask, if ANY NOVEL PHENOTYPES APPEAR.MCB 140 2/16/05 2014.11J. Lederberg, E. Tatum (1946)Novel genotypes in mixed cultures of biochemical mutants of bacteria. Cold Spring Harbor Symp. Quant. Biol. 11: 113-114.MCB 140 2/16/05 21ParameciumMCB 140 2/16/05 22B.H. and A.H.In the pre-Hayes period, mating in bacteria was envisioned as a conventional sex process, perhaps modified by aspects of “relative sexuality,” but nevertheless a standard haploid/diploid/meiosis mechanism. After Hayes, it was known that bacteria were not just small cells, but constituted a completely different kind of cell … The terms prokaryote and eukaryote were not introduced until 1962. Brock p. 87MCB 140 2/16/05 23William HayesMCB 140 2/16/05 24Selman WaksmanstreptomycinMCB 140 2/16/05 25Hayes expt:Take strain A, which is streptomycin-resistant, and auxotrophic for biotin and methionine.Take strain B, which is streptomycin-sensitive, and auxotrophic for threonine and leucine.Mix the two on a minimal-medium plate containing streptomycin.Wait and see.MCB 140 2/16/05 26Rich husband, poor wife is not the same as poor husband, rich wifeCross #1:Strain A (StrR, B-, M-) Strain B (StrS, L-, T-)Result: streptomycin completely inhibits prototroph formation (i.e., appearance of B+,M+,L+,T+ bacteria) if added before conjugation is complete.Cross #2:Strain A (StrS, B-, M-) Strain B (StrR, L-, T-)Result: streptomycin has no effect whatsoever. You can add it all you want, at any time, and prototrophs will still form!!MCB 140 2/16/05
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