por" Mepplrmintiny Meemrbmiane Sy bee 4 1447. See (ssa? VYYo x W-l. 7 Plate v. thle m ENB LH (6,). erde, EYSHLk £6, boots fox arcloumns - Conctnel, M(o). Vara Ba + i. 4a 32 o l 48 7 d 22 f v chiar ster Q) 1% nt © 22 «4 2 | 266 2. 4(6,). 13 5S tpl Lip Dez + See . t 7 3 2 ig «6 © 2? ppete frag bell lacr y % 6° wf lt ‘ a 2) oO % Bee APL 1 Lee MY, ¥ Vv, Aur Yee G1 JEL? Ye/o x Wo). ee oe veuplEn iA Sera by mtn Krontrnats — , [tad yi, fo). Malt Ml—- Sell Sun 2 Ye 2 268. 2. H4(6,) $5 VAS § 30S. seu. 31 £29 7 S72 {lan 067 92,50 072 J 20, 0 | About 8%, feces ~t Hable +. 3. bael). laet+ Laee- See. a 94 189 5S 293 ~ 4 328 (646 O17 ef) é Fim LC hee. Lact R LactS lak bar - Ere oe. Nal+. Y 12 IC os v Nok - 4S / 66 7 WP « LPO Ge yore. nlf, “SAD. oe Fann 2, Ce bee. 3 Malt Jo 6 16 cf Mal Yo tal | Melt Ro alt S Habl-R Mab -S. F tom 3.% lal. So pact. I 0 be 3 7O C Lee - O 2 £9 2) BQ, pate. SIA. Mattie & aan y a Play mer a Nellore +. 12 Lar - vy" Mallen + . Bs ogee Jo lLac- UV": 6 haere V,%. Mellerr +. 2r Lex - —_ fe ben th Mhas - lacksre f ' “4a * . Halt RW, | Mab -R . T / q oO te 6 4 oO & QO q 7 6S Te 6b. BS 70 ee” lee oO O02 BF af } 6 oO 6 3 / yf ] } 7 ° oO 2 o 3 IS 3 iy 9 3 é o SI. - YS, ( Q gy we * g lod arent 9a ~ S 2 a - ¥ = 1) DHYmM YA Oy ~~ CulMir Ae m nalbre - _ + 3 . = ’ 8 i, . t:- 272 1S nt : wyy yy 2yY 835: Zo. eel sa | Hes leet: - om, Stak +. 4: [Dlact+: Place — ; 72 +: (0 ~ 3S Lact 19 tee] SY fle aaa bapke conapaccd Co 4S, eagles ragels @ reap dedancce cx Cnet thel frond on the draees of Che bec, V ole. A lyse uo, He Mah Rigany rang fy lactase (6) tommimacle favor abldimeal teresnus & tiple Wt [1 Nee 4, 1497. EMG Melle plate bald fr taseed W- and TI a2 Aportincs cepb thw fob areth weft wcll hifoal Viunerer, Sriaheudt oncsuel, (1) colon Ca ih we Mins. CF At TS. (et 70 SZ [ren nece Cole oflcloes foerenlains 0-49 td) ~| 5 3_ dec. 7, rte 7 Wed 5 (Lac, - Pt Lace x bac, + By -bae- Malf~ ) wet. | plete debetien, 8 EMS lac nate sj} Fo. 68 | 192 ‘nalef fo, Cz, ‘ slich pctec miner thf 8 | pnrierrdle hae - gun fen _ og n. | proenela Le fev ctaresrebrhele | me oF, s “ tL 42 bac betel, BY) ML | Mal + Feat of of Hare Lae evcdibodely on baclpas 15 Mnelurm . in ame wr hear crerreslonns (eathy ¢ a, 4 hed 09 corereead Le Ugerd Har Lac, Lac, ~ . Shelagh jr EMP bar rset regs lll oly lk oS [tte park 53-6 —uste that Cire retonscs une um fap Leena’ hhist ! SY [7 ( Le Ay - Run, “ . ubelen, i - {2os0. Like, [pee = tHorcesr fj 8.« 6.) 15D jAracat te , , e Coote 54, 5. Lac. tee 1D reecld o@ Woe 13, 1947. Decke+ Lehl Meer clones este hth Gre lan 13. a - Wsb Gave t+ W-ST. Ma coe nigtt Reskiinly we T4~(5 Rewre. 64-16 Rew. 17 No aveadsel poration . 1P-20 fare’. 1€: wo (G: wel Wo wé2r- 22. Ar tl 23. @eo lf bo. 15, 997. sj) hewtecake - Aged SYAL. We Leff seat arotul | SY-/, 3 Gee util 7 wre C Apel Conger $ zt w 63 5Y~)f a) Mk He ») bbe, poctiy tical 15. Teruel Bly. Rectal Gee Yl bo Lhes. ahal at. 1h. Reatircele . 2) Keotiah 22, Re hiialh . 23 Tok . ty Woy Us Wst (Atol+ ). Vo Sakrs. Nich lb vlauf. 2 BE = - — We7 tel gg heathy wee + 67? oF wr? 2. i puretabl pack, Y, iat + and~ ~ wW7- t+ WPS 3. ttt tnd ~ ~ WY rs SY, pee 16, 4 74T, a+ W74 b.- W78 Ld - at Wal b.-nx wW8o + wees ? Depoet Prantl ban ¢ S-~ an + tatorrees . 5 uw ey b * Agente, — Carlos ‘ aa Lf there WPS / . ¢ a WIP 2 7 we P &® lac — tMhKhen ty) hae seis,’ Dee. F447. arhcahi, . Breede Tirana bac bianacs m specthaten’ ba C sch fe sdaed fin howe, Eopoed “capone fom pcos Rurruno Y¥53. + + + Lo345 _ sp + + & + + + at + w30 + + + + 4 4 + + + W % - a st a wo + t ot. 187. 4+ + bacwtt © Host +4. Sume Lae ~ a. bee +t €. Lar - b lac ++ 4. bac +4 h. bee — BP lac A. bec - Ts - & x 16 6 ~ | bb Lae c + + f S816] int + ++ w-35 = - ‘ a 4 2% Kup nn ahaa . rr ne 5 wey Se « We dh Bibra te / ° q: . | he: Do, ff b, 1457, Y(320 to Aegmene F/0) +: nor. PIS. ny Ae kor Ff po! 3, MEH ~ 4, EAA ~ y. MtHE AA ~ & He ~ 7 Yay ttt & V,ts - 9. wcHVTS = iO.» tat > ~ 4 Neat Cilacl Mind ?? ng acne, “4 WF-12,3 im: Vale TO) + ' > 3 ) = + . @. FAD t+ oe ~ 3. WA : ~T 4, Vv, Fe Sharhy nan C. i: ae W- j Ist Aig" nd Veron TIO) , 4 aff “(tes we.) erg sce wb A-fA. av ilo) boc, at 22-29" " | i hee, 2°" ( iafoce G wwler meth A I, Yay, - ae 2. Urts. - re. W-l am To) + ae te Le oy glia CAB (R- ednopel fur sharLohnr} TT =—-_ 3. do.+ mater roneree 16 fwd. ann 4. do. t pol lr fwnQ + 4 — YIB2> YO" ntenky . ; u~| So. YWA. Peparating easels BS, Sedum Wercler Lee vy 125k My 0+ Ha 28° Mit weBe aay 50D arte haa Shope, Arnel ectortace of 1S (by. Yalleee ecco mr retatern Ss) bee. S, IGY7, Tet the follrwons 7 brcltatse EB. bb. Lane. Nba + - 1G 6. the Nblsin — : Ww RHE Ww tHt wr+l + W6O wet wssé wtadan +f +H wér tHE LER Ly 6 pee bey wts Dee (P1947, wa" + w- 71 + w-78 + - w-8o 4+. + Uulilon, of YI9Z. SP dee. 17, 1947. To) +. Alt.) ALS, . I. Y. Gy. oe +h “ W2 Care 2s arte (as alrw 2 oe . +t “ than ay ted . 3. 00s %, . # - . yf. 0005 %o ~ “ YMA bar prarrtadunl ~ ok, 5. YWwA. .6% - rae ca. .f —,ol al ; 6. Wilerxe .5% & t 7 _ _ nal Geen Ha, Mupitlirn tghidigreler, (5. alin , ectalhioniz;: Tho). f. ~ 2. ¥.ey.I% 3. Uhre JS Y Wr tare S% 104, 1. &yT. 1, e@ 20k 2H, fl, / x. PLE. pd (fla, (ba. fll. Ceregpoale ] r4Ueacdt nnd jabae un B 2k Ht, 0. De. Kerestue Fela 59 Lac-l and Lac-2 mixtures Yec 16, 1947. dake up 10 mb tubes of lactose 1% BCP broth. Add .5 ml inocula of : Set up 2Pih 6 P16 10 a lp ALS x W-45 - tt W-45 ~ t+ W-54 - a We54 - The Same - WAS & W54 - t+ WAS & W54 - +t. K-12 Hy, K-12 On aun FwWDNE *herefore mixtures of Lac-l and Lac-2 are unable to utilize lactose, although recombimnts are able. ¥ steaalima my bacle rcbably procs Heath + colores Beals Botte yt w-45® lh bun, farts. tecombination of Lac-l and Lac-2 on complete EB medium 61 Yec. 17, 1947. Harvestxte W-45 (Malf Lac-1/ Lac-2—)aak and W~54(fal-Lac-l-Lac2/ vy," ) from fresh YP cultures, and mix at a conc. ca 1010 fml each in water. Store over night in refrigerator. Dilute to 103 / ml. and spread .1 ml on EMB-Lac (NZCase) plates to detect? possible Lacl/Lac2/ recombinants. 12/18 PM. 121 plates x 357/2 totalling ca. 40,000 colonies examined. None were Lac#, This is a control on the reversion of both Lac-l and Lac-2 The recombination rate under these sonditions is apparently too low, 62- Nutrition of Y132 Dez. 18, 1947. PIS Inoc. into T(A) 4 AIG. P19 har i. Ye Extr. 05% $44 - 2. Y. Extr. .05% ++ “ 3. YX Residue We vt — 4. YX Residue .Q5 rT LC _ 5. YX Extract g5% AK ihiwt 4 thea 6. YX Extract 42% - “ de Gelatin jvdiclyeste 5% + + 8. Tomat - a omato = ce p> +4 10. NZTone .3% Hy Lv 12. NZAmine B 05% tt t 13. YNA intact (NaNucl.) $4 * 14. YNA hydr. o§% | - -_ 15. YNA hydr. o§% # YX 05% test tit te for inhibition. Idfeodiiorinocterx 16. —_— - ” 17, M2taae Si + H+ C Mbnn or acti &) 18. CBee Cbe ard 001% oO. Fou a tality no nol deel Ly lbba sider feed spot) (Mgt U2Pase 2] 10g. Yeast Extract Yifco extracted with 40 ml. CHC13 in flask. Separate, evapotate extract and take up in water. Expressed in terms of original yeast content. (Very little material was extracted, perhaps 1~5mg. at most.) W/5-3 LY Plu) + Pry YX ,2Yo 4+ tt NeeV + rt Nigar - th. 63 2-Chloro~4—aminobenzoic acid Inhibtion and resistance mutations Dec. 15, 1947. Prepare plates of T(0) agar with 25 mg% CAB. Do. (0) agar. Spread ca 10“ cells of K-12 on both, incubate 72 hrs. A) T(o) agar: 400 colonies noted b) CAB: ca. 42 colonies noted. However, direct microscopic observation and smear impressions show a large number of "micro-colonies", probably equivalent to the difference between CAB and T90) plates. Each colony contains, as a guess 104 ~ 10? cells. This suggests that Strandskov's observations can be accounte d:for on the basis of spontaneous mutation and selection among the relatively large numbers of cells in the micro~colonies.) Maltose-segregation and supplementary genotypes 64 Dec. 16, 1947. Harvest from YP and cross W-55 x W-54, heterozygous for Lacy, Mal, Sal ,B) as well as V,". Cross on Eils-maltose with .002% glucose added. 4f,. A20. a) Estimate frequency of maltose‘, and of sectored colonies. Score only those where the sectors could be scored accurately. Proportion of Mal / (including sectors). Count sectors as 1 4 andl -. + Sec. - , A 2 57 Melt 2 THT 7 YE 2 f o IS] hak 51245417 5 3Y Pgs 8 Ss. 7 LDR. 12% | 17 Ss si2 Proporthon of sectored to plus colonies: (Score under conditions stated above) Phale See. ha # Jee tf See +. tb 2 j 5 ¥ O ’ 2. 3 © 6 ! uf | O 3 6 > f } / O tf 2 2?— | I { Yr O Ve a 3 0 I } ] / Y {1 d 3 ! a Loy 3 2 oy | 3 f 3 QO wv yf 2 2 2 t Uf 2 3 | 2 2 3 O tL QW 32. [20 (72. | ; 0 30% fhe Mehr colar y i y 2 3 4 2 vigo Harta Nal- Lesyesacd . } * //) : = { vv _ { 64A a20 Score Mal~ segregants re Lac and V1. Also Score Mal f Mal— Lect VU tae V> Car-0" fee -Y> (Vel scored well rn, i e) o,f, 1b 5,4 4 2 , , Ge lanes rertaomnal fo 4 cert mn wal £ bare v4 Lact lac -U" lae ~? “ty Recgue lt Heer ¢ fate, £4 S* ta aa fasta Pick 57 apparently sectored colonies to water N20. Store in refrigerdidor for laver separation. Hecale out 7 "tol 640 scores on ial p/m components of malgtose sectored colonies. Lae p/m and V, r/s Colony DMWNIOUWE WH PF kt .o idal p ms - ps: ms - ms. ps- ms. ps- fas: ms. mr. ps: ps’ mr. ms. ms- ps: ps. ms - Mal g Scoring very clear except where total lysis may have obscured fermentation reading in 22 ( Ysaxys5.) » Mab ~ lee, - v,* x Holt Los +V,— Lac, V, scores of intact colonies: A) Mal / ~ R -S +R + §. Ss uy oO 7 2z 7 O 9 3 uy oO /O 2 of o ' {2 /9 O BT / $8 24 “Rave Fa yf B)Mt- 5 7 | 10 7 $ ° 10 5 6 z S y 7 oO /o f 3 Q eM 15 28 3 H6 [1 = PO 8 A 37. 47 3 $Y _ 160 . . £23.21 Coal ¢€1.97) CV¥E.7I Cegieat wal Fille 5 Ahmed paps. Lr . gs = 26 ( ° Dart. ( Sf Aicclecs pops , i Her iceabue beden’ feu? Mek suk te bliin Marl B! (ne wbicecling with ba), Mibat. D7 v7 » 7> 160. Geld Yhy wey yu Go. Noles Mane F-R anid difresney of +5. A ng. M. bere bee / y, f- _T4 . i. a arth, b. ps 4 O ! “rg N+ [rt - a y, + Te = YZ . 30s Io>L+9$ +» 22 4 fejech a preter terhminand ub yak tay TH. The difference between large and small colony noted on the above survival plates breeds true on the first restreaking on EMB-Lac. Transfer to slants as 67~1 and 67-2 for the large and small respectivelgy. TREHALOSE MUTATION RUNS 68 Dec. 25, 1947. Use same cells of 58-161 as in Exp. 67. Expose .1 ml culture per plate to 15 secs. UV from GE AH-4 lamp, as in 67. EMO - Tretialore pater A). 9 plates. sdost plates, due to faulty pouring, had pitted surfaces and were very unsuitable for scoring. 1 plates, prepared earlier was more satisfactory. this had many (5-10%) colonies which had radial striae suggesting namerous variations affecting intensity of fermentation. Hold plate in refrigerator for later testing. i[3/77. Mo rate food) Maltose Mutation Run Dec. 25, 1947. “et up as in 68. EMB Mal plates. Ca. 11 x 200 or 2200 colonies. 1 sector noted. Pick to slant as 69-1 for later verification. = large and small colony types seen as above. Janr|3 Suehodl as EMS beatend ALL Hel. Mo uuctulas 69 Lactose siutation run Dec. 26, 1947. 2et up as 68. Lac-EMB plates. Killing variable. Ca. 200 scoreable survivors per plate averfage 65 plates, or ca. 13,600 colonies examined. 5 possible sectors. / II 4 core df 2 Wot sehr 3. (e No unrfauta _ ACL lace 4. OD taud-. 5 bee : ® Vo bec rtectrrete Maw Mecctore) . hiclile ato 219778. Shiba, ef Eris 70 wutations re beta-glycosides. 71 Dee 27 1947. Grow W-55 (58-161 salicin pos. mutat,) in YP broth, harvest and cone. to ca. 1010 ml. Irradiate 3 ml. in quartz flask ca 5 secs. rotating at font of Hanovia UV lamp, and inoc. ,5 ml samples into 50 ml T(m) with .05% sugar. 1. Unirr. a. salicing.14) 3f B. Cellobiose - C. b-e Glucoside 34/ <. 5. Cellobiose - C. beiie~Glucoside 37 Incubate 37° Examine Jan 3 48. ~treak out the b-wdeGl cultures on similar kam EvB plates. +his may be slow rather than mutative utilization ,a s has been observed before, 1 ea ie foe pe Piceee OE eth fon proces Mopar an AB8. & ) t ar tamrmtal yng a thas _ rs a Bear. ote 76 (oo) Pap facoe b colniepolelor, £0 genet. Als. 1B (Cbbtewin) wtp bene vechad 114, vlele bes LL) Al3 Fre anges tah acento f Keli’ | Kicbe Cross-lest sini tose mutants Jan 4 1948 6K. i-1l Xx Pesudele A8 (Ppoton # -“ — 1 W-56 - 2 w-58 + Ca tito Malt 3 ii-60 + ca 1:20 mee ey Trae” Pau ra 5 a-71 + Calico 6 W-78 ro ta 150 Mal t. oP ae Wee BO +t 8 we 20 + ca 1.50 wbx YSR ghht Parent +. weSornysz allt abw-l 4 W-S6. b) WS8 Wha W-20, W-7/, W-76. ait deed od Up 0 -_ + wl. OX, w ye. ok, wWw58 ox UW bo. On. w 7h OR. Nuyntigus Shieh nD 78: Mal ce Herel oe 73 trehalose sutation “un Th Jan. 5, 1948 Irradiate Y-53 on Tre-FiB plates, 10 sec. under Hanovia UV lam. ' 10 plates. 10x ca lS> ~ Foo eolorrced . Aeclowee's slipmed fork, heatincd Aeelovny t 2. (2 On > bit og if andr 7 ° fa ise, BP nd Alen b. ~ ‘\ untal options . ( fite’s Way le dua to ‘pda _ - ~ ~ me tw -70 yo- Fr Fee 79 tothe rhe sabe of age See. TS wet, OE teen “ dla ~ tok, pe ee pl Aik, ” . tratadrle ths vp cok. [he 7 nf” Cy" Wd vk “y 89 +4 Jo + ty ++ Ge. 34. RIGA pw. Tor EMO. ( Hlgy Tabretoas by f bias C~-l Fermentation “nutants 75 Jan 5, 1948. 3P. Inoc. Y105 into YP broth to obtain calls. —f ee eer Ure Ybuep " Quadeabs ey Cy en ifn op Oy Has at Le 2. Nw ENB 37pldis * e090 [pate . = 700 colnrers . ; Mo vexxr tamHe Leet 1h 39 x ta 26 = 7232 tlreees | | Jae eho ual steale nef ny Yo! and! Late. MNol—- on. Lact vad wiooplerleahle Aiviletal vl, Has "ypeer. 1. Re 36 4s 5. 6. 7. Nutrition of #93 Jane 9 1947 Inoc from fresh slant into: T(Val) plus: UPA. HC + Vits ~ HCVits ++ NZVits hep HCOV~ YNA _ XY. “xtr $td+ Sodichwaex 75 Looks fv sp, women