roe a Madison 4, \ ie ae ge oe 7 POE Dear Bruce: Your lettar of the 17th received yestarday, the me. a few days befors, but today waa the first chance I had te get to it. When I finally did, I enjoyed it thoroughly-- hope my amendation does not seem too abandoned. ‘The details are marked as far as I can on the mse. Some general pointe: Terminology: eazecialiy trameduction . Treated in situ. Punctuation: I admire your persistence, but I think there are too many semicolons. In general your own criticiom is well taken. The experimental parts are quite brisk, the introduction rather overworded. Fat has its uses, but I've suggested some places where it can be trinméd. As you say, I think have have brought in a number of side-iasues which not only take up space but may distract the reader from the main argument. Thks is as clear as could be, however. Do ygu wish to discuss authorship any further? I gather that after you decided not to assyme it to yourself, as I originally suggested, that I might still exercise my own option. I would still prefer the first pian, but recognize Norton's vested interest in vetoing it, and will in fact agree (after more prolonged judgment) that the triple authorehip 1s wiser. The advantages, of expedition snd simplicity, are already abandoned/ so there is no point in reviving my first proposal. I am sorry, in a way, to report that SW-553 also shows a linked transduction. (2/60 LT-2 -~x SW-967 (another isolate of 553) were i:--.)It wae originally detected on g.. serum agar, later with isolated swarms on straight semi-solid. Is it worth the trouble to bring that in here? I have no personal ebjectinngxbet predilection, one wey or the other. but suspect you might rebel at the idea of including anything new. Thee may be some sircumstantial evidence of a partial homology of SW-967 with SW-666. They transmotilize each other all right, but the ratio of tracks to wwarms seems rather higher than when, e.g., SW-666 Pla --x SW-967. SW-666 --x 967 is the one that shows .such tremendous tracka ( best followed over a period of two to three dave (sic) in tubes of semisolid inoculated at the surface. J would estimate the full length of some of these tracks at over 30 mm (over 2 om in projection), and several hundred microcolonies. No major branchings. LT2--x541 and --x 548 were reported as 1:1,2... I have about half ml. of absorbed 2 serum; 3 ie a difficult reagent and no longer recognized in the schene. Felix may be playing games. I've been corresponding with him on the same points} Boulgakoy phage, and stability of 0-901. I compared his own old isolates with our stocks, they look pretty much alike. Am sending SW-970, 972. The latter was lebelled aa "aberrant gallinarum" by Kauffmann They areftxks completely stable, with or without any FA trisd, but 970--x SW666 and 9%2--x666 have both given bi- and gesi-. They tronsduce motility to some Fle- but not others (3,5,8 and 4,5,5) and co may be tanzible exampler of stability by mutiple Pla-. The homology tests seem your business more than mine: want to try them? See what K says about SL-13! Edwards takes a dim view of this kind of analysis for the imstant purpose: the differences may be essentially quantitative. Pullorum strains labelled XII, XIIz and XII, XIIg seem equally susceptible to PLT22. But he did give me some durazzo and XIIo strains and serums which should give the necessary reagents if I can get around to the cross-absorptions. antigenic I am not so sure of differences between early and late swarms, but there are defi- nitely some much earlier than othera (the later taking 18-24 hours io. ' ar BLA ) Ara A At ag Sefoke » BE th LA AL [Lab, next day] I don't recommend saying anything at all about K-12 recombination, if it can be helped. The differences have been gone into into the previous papers where they are more relevant. The best way to avoid any confusion is not to mention K-12 at all. I am hoping that Nort' may pay us a visit in connection with Federation Meetings in Chicago second week of next mothh. It will be a chance to coordinate some of our misdirections, if you can return coment by then (if nesasgary). Good luck on your possible appodatwent. It im't, by any favorable chance, thie side of the “tlantic, 1s it? Do you know a chap named Aleck Bernstein? He has an Mel). and a Dipl. Bact. from your place, would like to work:for a Ph.D. here. JI think I'll probably take him, but would welcome any comment or advice. (My j-dgment of Efigiish hacteriologists may be colored by recont wonderful exneriences). The paragraphs that follow have been written at odd intervals, and may be discon- nected. I can cohfirm tracks and occasional swarms from pullorum—x 0-901 and gallinarum —x 0-901. In view of the apparently rather low efficiency of transduction in this system it may or may not be easy to make an adeqpate test of H, transduction. I hope you don't mind that I duplicate your attempts: as you will note from my Aast letter I have been engaged with this, but with rather negative results so far. I am especially suprised at having been unable to tranduce Mal+ from gallinarum to pullorum, but have now to try 4 Molt pullorum as donor. I suggest that a more explicit treatment be given of the behavior of 0-901. This is probably the most widely familiar O strain, and some readers are bound to try some experiments with it. I have not tried a complete homology test, but it might be sufficient to empkais emphasize that k®xdaaer 0-901 (including authentic strains from Felix) has¥ not only reverted spontaneously, but hes¢ given additional additional tracks and swarms, alld, x— LT-~2. In fact, it might be a good idea to use this as a way to emphasize the efficiency of the selective technique, quoting Felix (J Hyg 49:94) to show its stabilityf for 28 years in ordinary culture) and alsc to minimize any sense in which the discrepancy might be regarded as reflecting on F. When we first discussed this, I had not seen your paper and did not realize how much genetics you would be able to communicate. I leave to your judgment (as of course all the rest of these remarks; key the only thing I have any strong brief for is the rectification of transducticn-tramsformation) whether to add the paragraph at p. 35 justifying the concept of linkage. Hm I think if you introfluce this concept at all, and I admire tam your success in clarifying it at this level, that a little more should be said abet it. On the other hahd, I would not want to preempt a more extensive discussion” I hope we can elaborate at some future date when the 666- SL-13-SW553 relationships are further clarified, Edwards! report just in on the motilized forms of the following, being sent along with SW-970, 972. Here is a fuller listing: motLlized: reported SW Edwards serotype remarks 963 4936-50 4:1,2 ) 29 965 no good record i:~- monophasic slow, irregular spreqi "Zelly"~1950-Seattle These had been recorded simply as group B. They were motilized with x— LT-2; I will cheek using x— other serotype to be sure. 970 3821-52 D nonmotile gee not motilizedg but —-x SW666 971 5465-52 D nonmotile(Texas) gam:—(enteritidis) 972 1553-52 (= Kauffmann's 151-52) 2... not motilized but --x SW666 I will see whatever further information I can get, but the records are often fragmentary, and it 1a considerable trouble to Edwards to trace them back. iIf they had the money they could use an IBM system! May I retain the tables? T can indicate my remarks here as well as on the sheets. Table 1: a more explicit designation of LT~2 (which is only thse phage type) is Lilleengen # 85 (isolated in 1946 from an autbreak at Stoke Mandeville—— Lillengen p-68 JT may know more ahopt this.) I am heginning to find LT-2 somewhat awkward, and would second (but not initiate)a suggestion to xkama rename it TMl or something of the sort(TM2?). This would at least already carry the connotation of typhimurium in, a.g., TM2 —X.... If we don't alter it now, we'll be stuck with it forever; it would still be time at present, and the equivalence could be put in the table. Add 0-901 to the table, and let us adopt Felix! oldest transfer as our type. Have you been able to track down, in vrint, the exnlictt history of 0901? The only paper I haven't looked at yet (library trouble) is Felix 1930, Lancet. SW534: add Sertic—Boulgakov refernce for VITI-133 . 533: Edwards #157 (N25 isxmm a lab. designation.) There is a referance to this, perhaps not essential here, Edwards & Bruner J. Ract 52:494 (though Cherry actually did this). I have not seen the phenomenon again, having always gotten 2.. rather than 1,2. It is not absolutelgy excludddyi (Cherry is ndt too clear about it) that the three occurrences were progeny of a single mtation. Other monophasic —-1,2 have behaved as Flap alleles. Xdunaowhkatx mx LI've told you about finding an old stock culture labelled SW533 which was a mixture of the diphasic resembling SW703 in ite fermentations and the —12 monophasic resembling #157 ] tithx SW541 : I quote ixockektaexx Kauffmann's letter (2/31.62) "Wr. 13 = §. typhimurium 3173 " SW544 "Nr. 58 = 8. typhi 0 901 7 542 "Nr. 223 = S. typhiomrium var. copenbhgen 1810 SW541 " Nr. 248 = S. paratyphi BO 19" SW 543 Edwards' 13 has the same description as Kauffmann's. Ditto 58. They must have a common list (up to a point). mendelian Add: “ ‘¢ SW552. Have you been able to do anything with this particular isolate? It has been implacably rough to me. Its histroy as far as known is on the enclosed photostat. I have not yet had a reply from Davila. All the Salmonellas indicated are gp:- TABLE 3a. For typography, I suggest i:1... for diphasic forms, The third column is redundant, and could be implied in heading or footing to the table, This could save considerable space by permitting it to be set up in a single column width. Id. for columns 2-3-4 of 3b. ‘ TABLE 2. Cultures reported as TM by Edwards are i:1,2... (I must have mentioned this supra.) What are your frequencies of a,i in TM2 (sic)—-x SL13. As I recall, I found 2:2. Tt might be worth mentioning (if you concur) the very low ylelds in this transduction, You've gotten a motile from SW573 in two steps, haven't you? I would put'8pontaneous" before mutation in heading, column 4. SW534Fla* is also 1,2... Is table 5 necessary? The data could be given more compactly in text, where they are essentially repeated maywhkex anyhow. fable 4. Is effect of SL18 oh 573 consistent? rather tak + S and F as symbols, why not define + as fast swarms, and omit F as symbol. Inset, page 25 and innumerable factors controlling chkoroplast development in maize, of which 73 were already catalogued twenty years ago(de Haan,H. 1933 Inheritance of chlorophyll deficiencies. Bibliographica Genetica, 10: 357—- 416). Hal You could also quote many fecters affecting the tail in mice (Gruneberg). There does not seem to be a more recent cataloguing of plastid effects. This happend to be the most cpmmon type of mtant seen in most plants. I just remembered a reference you should quote: Lewin on nonflegellated and pazalyzed CH domonas: JGM 6:233 1952, esp. 242. I have been trying to stimulate further his already active awareness of serological possibilities. ACKNOWLEDGMENTS sees. The work at Madison has been supported by [fellowship to B.A.D.S. [7] ~ from the Commonwealth Fund and by] grahts Siemmcthundiatiamrkxtk (E72-C4) frog _ the National Microbiological Institute of the National Institute of Health, ' U. S. Public Health Service, frpm the Rockefeller Foundation, and from the ' Ybbeemmurioctkx Research Committee of the Graduate School from funds supplied by the Wisconsin Alumni Research Foundation. This paper is recorded as No.536 from the Department of Genetics, Agricultural Experiment Station, University of Wisconsin. We are indebted to ...Kaufffmann, Felix, Edwards, Taylor, Boulgakov, Lilleengen, Leifson for generously providing cultures and other materials. 35 inset ] uv . The genetic,analysis of the linked transductions is being stucteti- tau) further, but two lines of evidence may be quoted in support of this inter— pretation. Yhmxahbxkity-—to-sive—-swapmas-whose-flasellLan-antigsas The two 0 strains, Si 543 and SL 13 have been exposed to lysates of numerous other types in addition to S. typhi-murium, and in each case where swarms were produced a minority of these conférmed to the Rake flagellar type of the donor strain. Thus, in addition to the stotype IV V XII b:—, SW 543 has engendered other monophasic typesf such as IV V XII a:--, IV ¥ XII c:-~, and IV V XII on: — when exposed to xXyamkssx lysates of S. sendai, S. altendorf and hedelber 5. ASOT respectively. Similar}y, SL 13 produced, in addition to I II XII ai—, swarms of the type I II XII r:-- when exposed to lysates of s. heidelberg. As some of these types have never been reported previously, their present exis- tence provides whatever testimony may be needed to exclude the possibility of contamination in these experiments. Some of the monophasic types produced in this way may be expected to provide antigens useful in the preparation of diagnostic 4 antisera#y (cf. Edwards and Bruner '49). te toe topeo of ~ cited experiments where 3wW 543 gave rise bo bg ant wee + erp e swarms 5 ete. coke mg X be thowyht 8 > each single. 8 5. ky - eee . aw. the transduction of ‘mm, cifferent sdinmbe ened fac t rather than of two a ray linked factors in the latter case. However, lysates prepared from such i:-- derivate Swarms were again able to evoke both B:—- and i:-— from Sil-543, and—the-scame—baekeres-st+ a dered . The ability of the i:-- A derived_froen ciefA2, again to evoke two sxanuduetiva types from its parent 3/543 is given as evidence that thezitam had received two elements from S. typhimurium, as distinguished above. Section I Introduction: The Phenomenon of transduction (i 15 2, k, A 2st. s1) Hereditary properties can be transferred or transduced frog one Balmobella strain to another by means of cell-free culture filtrates. —To demoastrate__transducien , a_donor Salmonella was lysed by a suitable phage (or otherwise treated to the same effect), and the lysate filtered. sxxkeokmixtax The filtrates contained an agent which) man, ‘transfetred various cultural characteristics from the donor mkxakm to a smal propor tion of sum exposed recipient cells/ of another strain. As a rule, a maximm per pillion of one recipient cells was altered, sven under optimum conditions. Cells which had thus acquired a new characteristic by transduction transmitted dnkx it through an unlimited number of generations, se far as has been tested. peer treseit | Hereditary changes, so-called transfomations, have been described in the pneumococcus ,and in Haemophilus influenxa under the influence of cell-free lysa&es of other strains, a ane eo recognized as being also based on the transfer of Neem genetic factors, transduction may occur commonly among bacteria. In these organisms, however, the active principle depends on the content of polymerized desoxyribonucleic acids in the lystaes, and its activity is desroyed by exposure to desoxyribonuclease( . ). The transducing activity of Salmonella lysates is, however, insueseptible to this enzyme, and is apparently associated with phage particles themselves (Z&L 52) Bruce: I have suggested this recast for two reasons: 1) the distinction between transduction and transformation was improperly drawn{ I regard transformation as too vague a term to dignify so important a phenomenon, but these particular instances af "¢ransformation# are indubitably transductions, as defined in Z&L. Many other "transformations" in bacteriology are equally certainly nct. 2) I think you may have repeated téo much evidentiary detail from Z&L— the reader get this directly, if necessary, and the conclusions are better emphasized