ML DATE: Pi 0l Jost pyrrnda F prtic Blac @D /koge tpbte . | Cypeer T Momre far [Pare, 7M elle ~ (2) wh in pasbch , AiR abrabo~ Blox. ¥//2 Raathentrep| Lee 10IP1B. Gare dae rp peel RL hom (pbs o Ralfeduel|—Bhe. O/C By ht. urke~to tered shee ppote. afr. ew? 240 L WwBed t ode : NIV—3 ke gant Miq-lxo geh™ | Stole rmnichate sails of. 20 30 40 50 9/10 /s-b ewe fos PL bpeoi _ Wo 1AM. fork. lof AW latinte om poet As fol. LAA me wotehin,| (ere |Rermtend poperr) - pth Dr eieby Kad * x Lv 265-7 wes 4 wo i4 sel w3077 wht S | welt 10] W246 4 wihSS wl W30G7 8 es | Ww7oiS wZHO W3/59 HE iv 3010 w313G — |WwReI77 Aen. Aah -}h Pl he (AARD- O Nase tetny dn. Fe F por - TPM > (og PU. PrgeTto Spa hy Asem Ted nce rote Hy - he pte bated) oC Reeneel fan ylittly, SK ober deen of KEIO | o/ Ve Hepeteny PA vo tn 30 [eo Xiao x iro K 10 = /d ye AAD. — ane | gy ML RIES SHE, ae Rea f- LAP x a ir f bs > - 7G “ 2. RE (a 16 25 weal ne Thy U6 Rae , SR | Pe FI pe ne et Jo~ tol lyeol 46 oe a posytercell, i zory meter jy o® Pl /ne other hea . Ww 1&5 ex fol TE SAL Tl bye net got | gg TAR TG Ayelet Agere IEG (VET). OT Tb parts : hs pak | lt ale 7 TS pref. He Neen oman of fns, PL Cymee - citer | obrrneet ee pot oth str . — DATE: VHe1SE REF: Wiecae ee 3 4 s whee rT TLBG, lo | F aren pARe col. Se ae . 10 rr Lg (. Ino 4 LM tl. WP// 2. |e T/1B6 fanadt ss — t NH s Cc 20 YL Dace] L L nn 71S hart aren | Molt prO, — PY/E fonth rire Bely ~ WIT Spot ont D0) + prctne bb repre TG | (Of perpen, 2 yee > lo) de) B Sae 20 BC faci) LOK TE Lor TT | 2 et\7/ 5 che Akira |areecof fiir taet mer, NT). Saree oe ~ Bie. STi de 2s - Be 50] bw /% 9 KI 15-45 | ma i ‘ ( S ) ~ N24 oar Led: REF: 1 2 3 4 5 6 7 8 9 10 o waar 10 1366 ptebesps a. 20 Wale aire 1230 | bby feng es . | TG? gone sed, TE wd 16 (urls). | Ved pas a) of -{ e Mm s } e ~| (. = 30 wilt FS Angra. Wits > RRL leyana, Wwe Joye B/I, Wwite > Tate of Tl, | Sor pna tn ene apd TE It Ft sol TE po " > oS —— So N27) SKA uel DATE: 7 [2 Gb [sb | REF: P Lye ote rn V3 dere > P) he Pi (word) 0 oO | TH lises) T/ (8/6) Té (4eS) 76 (B71) T& (HFS) Em. oY . oO oO Piling <) Oo 20 30 40 50 no oe om ley N25 tek (AT! L gogee /piXe Dm. LS Top Sons, payer + ohne coat, 4 i dma PI, | wigs + V1 Me Ut | (eee Heel) P| 3 Wi3GG + Pl de m wih G+(w 301) P| oR eR, (So | wer ees— + Pf Ae —— } 6 Mh EEF Cusvg) Pl [rnk. Pt Mpc (w 3014) 201 3am C3014) FL ma phe Mato . Arveard- Pike | Capo pete See SS _ 145k = GDR Fr LR D . Diseref- Spm) add tmd dL rn t pb.) ¢ o- : Bey Aseath, Adagtnny pte ce brg Gadd wne LX Al pets $0 [~Ase : q/sls& Toro We. ApS ves (@) Cl (doney) +H + Pl (oak deem be. es J ) O Pi (decprrant bE) Oo Ne 50 TE openers * boue~ Tee 00) vcr, T6( Bh) N2G DATE: 1 2 3 4 5 6 7 8 9 10 elite | Te le oe te w322f + |w3I33) Lo we | >. weep + | W3I20) + |w3230) +. | —— po, W0eR) 34D, 4 | w3B4¢| 4 feed) (ey) ae) rol gSo! fo | wRIr7 Sates eto, wws2l +e bv 195“) 4 w35¢ + WIEe i she. we 3174. + YR Te? | + 3 + wore of [eBnZjl + wr | ratte 37 veel — RD) aol TH + wea) +4 we¢o | + fw 2zzey, 3237) +. — w3/27 + w3240) 4 | — — w2e + pha — pR3& i” we 312 | 3234 30) Ww itZ 4O wT WAG QD w~WOF wigs W325 bo TU M24 VL I47 | wd 40 | 50 REF: - ye] Oo 10 FRM f 10 —lwea27 | 7 312.0, 343 iv 40 IKE Y b/94 “74S /7¢ 34K CFL (W333) witso| wes |) Be Tes 2D} ae B57 TN tel Uf Wel 8S wW/FY LS WAFEF w3lyh) ¢ s ¢ WISP VL7TA- : “SDATE: REF: 7 1 2 3 a 6 7 6\B 2 9 10 poe | wn BTS ES gH gee pe Ee be wy © O + O MF KF +t it wag) OQ t+ Ae 4r H+ tPF ik & | wrizepwr?2Zy pQkeT — oO 3139S [WZ wo W224 | w2UtS™ | WZ 2IP W323 9 @ 3237 +t + ++] | KOH te D 33440 0 |O +7/1 0 + + 1 & ae , pape |e TERA) eet) pater wef | WG | weil | TE whi) | TG W313 + Ww 3069 | te w7IS3 |) - W3230) + wr/Ge| W323 7 + (0) W3/34) +- w/S2.| w324O Of | ae ar7 274) win | HO) wre + w 3h Ww 2243 +/0} Wt w3ISF | 7S Fe | Re 22yy + fo] |) | Pte (2 ea Fh | an + W347 ee ot Jahres _ _ 313.3 | > W912 ie Hy W5io + WwI/5h lyp323B 32157] ~ . w3231 0 Ww31S-/ ARE) A 617 wT Z| Ww 3/93 wo 7 — [|W 21SG wasp] te |e3237 W366 | A pret 50 Uf °* H9/59| pe | WZ 2Y0 50 PRK oF B dinero bu FOLF OT ane ple 225- YRS ytraee woe) + 422434 | PRKE Io MTL wIs2, + ayy peti | Bigak- WB. Tom I MiA-Z Math REF: — TD 10 10 _ 20 ra "30 | wars> _|lwarro wry] +40) 50 WRCEq w lt & tv 34S D_ v[aals ws 7. (wats e. we BETA |w317s_ (we ses 3 | les2yol fy 243) ala, pee | (Ve e3 7) wR w 3174+ wrISe| W323 7) W3ne wrS 6 wos WII > w3Z2to w > [Qe by 2EH3 by 2 Uys 23 it f \ t+ tet tt | (4 eX ; Are Ta nee try 22 seer ol yt przy 4 @ “eo /e@ ®eeea ‘ e @-@)> “@ e cee e \ @6>0® 2. J 2 3 eo oe OY > > oo ° —~w N2§ Fat Mi3-BS opeineY wnurne Yared nM dre 4 BE for Lat vere, 1/26 (5E ner AI P Wo. j. | Wray’ | waz wi w BBP W323 | W2z 4} wad ite | t+ 44+ Af | t+ | et weeds) 4+ + + + + + L walt rei + ~ O + 7 © 10 w323e/t4 | + ol + + + + W323q +4 + Q + Oo oO wt +e | + 0 + | 0 7 2. *, plete 2 | | 88 W323) W334 [WIE T [WWTE WITT WrFY InBae 3 | SOUS HK YE 3 ~ ws 2 | 7 | © lel 12 4 s¥3 OO |) T Bt 1+ © (oO Ppa 3) G | /0O. JD I te | te LO 1 O sl | peak + W32MO! WIE | Lars W374 esi 6 WIT W353 Ww 3237 IMAB=2) 2 zai 3 2 2 | O G v3BF 2 2 | 2 | 4 | 4 9? 6 tale) CO | Oo |). o ! | m Oo 42335 {0 | 10 | 10/0 | 4 2 17 1 40}, FHKE Gw3W wr2sD [oy WIZT [WRHS [WV IBBF W323] HIT wee SF wt 3S Bil+ | CO ++ (7 way f | +H SF Blt) Oo rr) ae 343, D | FF O Bilt) o Ft) ge nes 10) Fr IO) JO]+ | + +7 /O sol] ae | 2 pr wrist 24 pate: Ww 3236 C7 NLA-MT XNE-L nye F agef— TLE, 1 c 2 1. 3 . 4 ; FN S L 7 8 9 10 seer ger (HER?) PP 27 A N6-t ret: p p ehmge Salen of w 3236, r-b Ph | SH Sg ekt MAG, AR Acta. ow Meal Sppten 6 lhe, GaRt deel a Sempre Cpeee rede aw VID an 4 bc. 10 Ons oka N29-G potsel ma Un KW CH t+ >), S Tike] o~ Birk br aye ote pdr setrLor~ . N 194-2 b4Leo predmadk | (4 62/2). Jolitfeb.| TA br Dio), Plo) Af | olis [56 Box 21-2. e22- Goreme tes | eg Pinty ; 2029 Ba A atr Bt ws 22]. Pry pol wadinly” w29-G. ( poles [5-6 wris3 Y10 w3oFF RYDE + © 7? 24-61 O a Oo Le w 146) - O t + 01 E156. Re KSaN” oan are 2742, 2776. DP-z 27-k |). WHR OK. = M . \oo eke Se & ee Wk |. >} - wwe, \- We7e, WR + Ww3I79st AH + Wais3 | 4 NX. E. orton Report on lac— study Oct. 4, 1956 Preparation of stocks The plan of this study is to prepare pairs of stocks cohtaining the same lac- allele, the initial member to carry Cavalli's Hfr, M, and a UV-induced lac-, the other to be a lac- F= prototroph derived from the first by recombination with Y10. For chromosome mapping, each Hfr stock will be modified by selection of Ves a marker closely linked on the left of lac-l, In future, the Hfr stock will also carry Pe, one locus for which is reported to lie between lac-l and Ve (Fried, Woe data fit equally well the order P_ V, _lac-l). an Hfr Pe be atock was obtained by UV irradiation of W1L&95 and is being tested to determine the location of P-. Preliminary tests indicate the order P vy lac or VE lac P, Pending the development of the Pe stock, F~ prototrophs and Hfr y53 wll2 lac? M- Pf stocks were prepared far the genes lacl®’, lacy ; jac, 7 2? 316 and for 12 lac derivatives of W195 (1940-51). wo7 lec, ; 1a W929 lacy In addition, F- or Fé prototrorhs were prepared for lacy”, lacy, lac, 193, w3128 and lac (Table 1 and Fig. 1). A. whe In the course of this work, we lac- stocks were isolated which differed in recombination and reversion patterns from the lac- parent:, W31L59 is a stable isolate from a cross of Y10 with the very highly mutable W1951, and fails to tecombine with W1951 and all but one of the apparently single-step lac-1 mutants. W3229 is a spontaneous derivative of W3120 accidentally isolated in serial transfer. It is much more stable than its 1acy®? ancestars and fails to recombine with any of the recognized lac-1 mutants. At present it is the means by which lac-1 is identified, since the lac-l pseudoalleles have sufficiently high recombination ratés to be indistinguishable from unlinked loci in streak tests. W3146 was isolated from a cross of ouil2 W3129 by W112 in a attempt to introduce lac, into an Hfr stock; it recombines with W112 and all tested lac-1 mutants and is almost certainly 2 not a derivative of W112, since it remains Ss. gale VE like W3129,. (Of the stocks in table 1, the Hfr tac? is the only one not yet prepared.) The origin of the two-step mutants W3229 and W3159 raises questions about the nature and frequency of spontaneous changes in recombinatio pattern of lac- mutants. Streak allelism tests Cross-streaks of Hfr M- lace end F~ lac- prototrophs on M lac plates are convenisant tests for allelism, but their interpretation, although elear in most cases, is in others made difficult by too frequent lacfé reversions, especially when they occur in the M~ line, and by the relatively bw fertility of 3H3, W3164, and W3140. Tests with highly fertile Hfr stocks have been unambiguous. The lac- stocks tested fall into two groups. The majority fail to recombine with W3229, and ere therefore designated lac-1 (Table 2). Of these Y87, Y53, W1950, and W1951 appear to be allelic, but may be separated by their reversion rates, which are in the order Y53< Y87< W1950 = W1951 when compared as prototrophs. The latter two stocks are exceptionally revertible and are probably identical, as they were isolated in the same experiment. Similarly, W1948 and W1949 have not been distinguished by recombination and revertibility tests. All other apparently single-step lac-1 mutants recombine with one another. Five lace genes remain unclassified with respect to locus, since they recombine with lech *”, lace2, 3,455, Fp ahd tac 7128 as well as with each other. The two recently obtained lac~ from W323%6 have not been adequately tested. With chromosome mapping tests, some of these unclassified genes will probably be found to be pgeadoallelic with known loci. Intensive allelism tests Quantitative recombination tests have been deferred until ve P- stocks are available. A few intensive allelism tests were carried out on material at hand, without re-isolation of stocks, so that reversions -~3- in the agar stabs over varying tire intervals were confounded with unavoidable reversions in the Penassay broths in which the cultures were grown up and on the M lac plates on which they were tested. Colonies were counted at 21, hrs. to minimize reversions on the plates. Despite the erudeness of these tests, they are of interest in confirming the cross-streak tests and providing a rough measure of reversion rates (Table 3). W3128 lace Hist. FY This stock was received from Borek as a questionable double mutant. Hist “ reversions on D(O) remain lac-. Lac» prototrophs were obtained from a cross with W1695. Both hist- and hist # were isolated from lacf/ reversions on B lac. All the evidence is consistent with independent origin of hist- and lac-, with hist¢ reversions in some lacf papillae. Persistent diplofids From Hl lacy 93 colonies were isolated which carried Het, as shown by laev colonies in the cross with W1940. The lac- parents have been stabbed as N13-2 and the jaev diploids as N13-1. An attempt was made to test allelism of the lac- segregants of H271, a diploid lacf which segregates stable and mutable lace. The original constitution of this stock was rad 3/1ac"*, which was lac- in phenotype. Unfortunately, the y53 Hfr tester is of low fertility and the wil2 tester iN oN Wil2 Y5SS wieg wos WHS = Wi33 W4s WG Z werd W JIN | W323G W3224 Source Moye y53 Vv wll2 “ wi941 v w1945 ~ w1946 wl948 ~~ wl949 w1950 w1951 v’ w3lZ6 w3159 w3229 ~ whd wl08 wo7 wi45 w133 w3128 w1940 wl942 wl943 wl944 wl947 w3237 w3240 Boe oe oe oe pe op op H NIow FF FP BW NY HE BP HE Table 1 Hir_ M ca W3120 - Lac Stocks 3H3 ind. Hfr (JL) W3221 ~M+ W1941 W1945 W1946 W1948 W1949 W1950 W1951 W3146 gal5 We Ss NO W3229 W3164 s™ W3140 Ss” Hfr M- Pa W3237 W3240 N6é NA oO Fe prototroph W3230 W3134 W3089 mal W3148 W3152 ‘1 7 W3153 W174: W3156 W3157 W3158 W3175 WE W3159 W3133 W3112 W2243 W2244 Ff W3127 : W2245 FY W3238 o ¥9239 FY W347 WwaZ9 * W3215 W151 " W3154 N23 N2 N9 N23 N7 Table 2, lac, recombination pattern Stocks recombine to give lacf if the corresponding bard do not overlap. 3ir¢ 2134 3191 3ue 987, y53, wigsnjy «2222 BBY 51571, 2156 3084 wil2 eT w1941 ai wl945 2 w3146 3175_ . 35 wl946 ° DID 31S3 3/33 3)33 w3229 Table 3 Allelism tests Exper. 1. O.1 ml. Fe snd 0,1 ml. Hfr from overnight cultures into penassay. After 4 brs. plate 0.1 of mix on M lac. F Hfr Me W3229(3H3 W1941 W3133 0 0 0 W3134 22 23 > 1000 W3148 1 13 0 W3029 oO — > 1000 ‘ Exper. 2. Mix centrifuged, washed with saline, concentrated in saline 1/10. 1.0 ml. of concentrate on M lac. W3229 W1941 W3133 3 2 W3089 0 = Exper. 3. 0.1 ml. Fe and 0.1 ml, Hfr in 10 ml. penassay, After 3 hrs. plate O.1 ml. on M lac. F- no Hfr allelic Hfr Hfr = W3229 W3133 0 W3229 0 0 W3134 44 3H3 52 50 W3089 0 _ _ 0 W348 0 wi941 0 0 W352 0 W1945 0 0 W3153 1, W1946 15 an W3174, 0 w1948 0 0 W3156 0 w1949 0 0 W3157 26400 W1950 14200 15400 W3158 29000 wi951 17000 23200 W359 0 W1951 32 0 W3175 0 W316 0 0 Table 3 (cont.) Exper. he 0.1 ml. Fe and 6.1 mi, Hfr in 10 ml. Penassaye After 24 hrs. plate 0.1 ml. on M lac. ¥F no Hfr allelic Hfr W3127 32 W3140 153 W3112 0 W3164 0 W3151 0 WL9A4 0 W3154 1 W1947 1 W3147 3 W1940 2 W3149 1 W1942 0 W3150 1 7 0 W3155 0 " 0 N30 Joe nr ! fon N 13-2. X ohare REF: N3 | 1 DATE: ef {( 5b 3 WwW 20 ~ 30 40 50 3iy2 SI] WIth Zam ws 27 3 N3a frtune Tet -f W3236 XW7YES- DATE: fo lESG REF: ~3236= M7 AFR. PO wIY¢S- = Gote Le, (7<28)- iO AW eye A prem P Ke, xX /o- “e~ De) t pork. loftlse@. She mm LP fer ame curQund 7 Arka. el atten Ppt Bec, lod Fhe My Be¥ otto. otk [. | Pret — | | Pam Pt] ee peta / /7 / hil tf peto2 23) YU et dt tt | fo! 3 | é |/7 °7o |623.5 | +8 GS | ARAL gt fee OP | Act fhm |2e/yB fp prise 17/43 : | _ 40426) boct ppl F b 5 V \ Be 4 _ $F 6d ood. _ hte) eT | epeot te frrro te T., 50] NaZA- a) 1 2 3 4 5 6 7 8 9 10 4 -—— CZ 7 Ger Pol | KB | 2 20-6 Ly | | Perf Vel be bbbe, DD AeZH YP = 70. ~~ AHL 1S YP & YS OF ° D BQ 222475 Hd 7 a) V-EPY-3= 206 E,W ory YF PL SEZ — 20 — 3 Ae He gah yo ~ AL) ¢-S1- + F DAL | Aer |? | CALLE 2 + # Eo ZT ur beet UP dnc | = ATH $ LIA = Az FIL) dae™ | Pmt + e* pe S747 APS" 74LE 20.6 “e ge VG dae. hen. mt fsa + - J ! Spor ee 2H] oh? 4-3 ~ Arabteg ©? haptic tit € oO Coin fly gee aris : /6 /3 N33 See vate ol] [s% K Vi resel Ine 3236 REF: — Fo pnrepte Urn af w>LBG reed prom Piece paren « Laren 0 inner t— tnlTerreos yr ' ya toe. UY (/OHere.s 4 Vi. W326 N33-1 foe 10 W3267 N334 22, —_ - 50 Wry DATE: T pane a rng rat. wt. Pl he REF: 1 2 3 4 5 6 7 8 9 10 pole (C8 Vb 4236 ome Shee He Oh ml. PI (u 1366): cohen lof b lus 236, 4m plli3eé), poLihhs & | Ww 3234 hr, to Pl 36) | dba d 4 | 3236 HPI (UII) Ca YOO py Hp ro plbe . S eek ATT Hore oo TET Pewee aL M&et Y/O aa hae S | Lec > 20) vrei | Co Reet OA te t+ Lng te “ est S let M+ Th oe t pat f- Ae Cad At? Wrrtedntal [MA oe : ~ lof 6/6’: Paspoa A ‘dn Blec.- Collel 76 fir fbr. 0 -Pnl 32-3 6 | ondnal P/(~}3 66) orf ~329h +4 fb Pllwireb) Repeat ~~ Stre+V| bee ea onS Ae 30 J 1 eSTeHn€ jy 32-316 ty tg 10 hee 7 2 & 324: . Ke OS ee P! (w i366) a BQ CH PL Mure cept fwd be WIEBE tH Cen 2ogertprite) where Apr ja [1U\ FE atrebhscl a~Y 1b wn M dee pects | HAE. ent Nee ee meme 40 4 & Z ~ sf Pl(w30e9) Je— ID Mt Cath AR on WI a ba. 1366) hangin pA On We 3s/27T a MeC- = uaew3 140 - ee Par bem BER? eee 30 ~/ - Spb3/27 ka ae ely y- 2 Cuser) Pl Goat 227366.) fl ome < “o.|K. i~3 “YSImD B/27K PS G2) 2-3 sPapgeer nla, 2er V3 + +) pels . 10 10/ (oe 10], 20) — RpRO Rey _ bef 40 50 Oo y ate tt + ett + of tt —$ 7 so+/ \ 0./¢. ) ety, Cpr) attelo~ Mae. Lrs3 W3)20, WI97 50 av pate: _jo//¢ [sc REF: Jo ore UY (idtp): id pierre of Lhe, (2 ef witso, lo{wa2zaG jo [18 priv inenn ee we LSA) lw BIZO nly pg be at Ae he Bet. [2 Lipa og” arabe . WolTP | Orra| Roe tr-—~ wi2>65V3 6-/er Kalil — Bee * TT “rn i rN ok Soe AT Xe> W3\r0 WII SD oxo 36-/ (w3089 36th ~4 ise | BORA | 56 | 2 wwe, Oo [=e | © O oe 23. |a32, O |e | O Oo O | ¢ luseg O |e") OO O O ~ S iw3im| CO | &B > © & & | wexws3, Oo | + O oOo | oO T 10389) OS | Blo O | 10/2))- Pr otal (2 pope 3i20 ) 2 bpm eit $0) so oo el Ong Abe che ~~ BIS en ix | 18 We | 2A | 28 Yio | RH) KE LK er IE w333| O |F |O | Oo |O was, 2. FILO | O © W3is3| 2 Fr), O 1|CO [O ” Taree A ~2 6B py, pofa3 Kier |e | 5 Gd othe Jom Aah Rs oo BO - Se eet Sees fA-| == 3267 /2C|—= 3270 ~ 24+ = 327/ 2B = 3272 N3°/ Wis66 Bek YT” TLE KW3Z36 YL PO DATE: [06 lI 4 ISG REF: val 1 3 4 5 6 7 8 9 10 Vo f( EASE. pun , page ccpatily wed) tute ~forgte tenis O/C. lofty] lke frtha Dor pork. SV oreo for Up, & L pho yhT, to fee I 10 (2.4) area ond yt crfernro on Bo, e/a Pra “hepa | Dat perk. | 0[2> FH} fh. Rapetln GO, Ble? B-o+T/, Bat7bs ~ 20 "30 ao __50 N47 k be—-N3 7. Chem peel tas of Tati 200 claato ules ante DATE: jolZ9 REF: fac | Par&® 10 OM MOMMA YAN \hyvt syyyad A addled saa tantaddsaaquistd Ud tddd daqadsdigadt | Jyh) anaeneee rates +p tel yp tydit pee etetH ll +) pd4+) 94411 ++) prtisett bittidgitidl peligf¢yrh dl odg Ye DEED LEE EEL EEA ERAALEA ELLE RKEAAL yatthadet tat a~ B-o+Tl, 76. Ss s\ \via\ \sA\\\iy \\ \M\ Wy Ww \\\\\ \\)\ Leaatktaat Latddddtar sadbadad 42494944 4948 Pan tly e deb laebb aid 41444) 44d 4 PisL eee tet t+ pypeeligie c dgebi dian rditaagp led Peidhe ra eid ete tpdtt it | aka dt heath Ladbhaaedtd beh Le ~t ptete, no Ve A fete an dake rtatat Laddtjttdd o we 5 an wy CV eh RBA y Neve. . g- oy ) 40 50 N378 REF: DATE: Wyn WY wd Muy aaatavcal TN dtdddaJ J SUH adrettadady 9st Qi glel pap Pe Heth yy pH ret Phixsdpyt ieee ld res apergibeg eet wea bate Eee eS tad eddy TTT \\\\ya Wi \Q\WQX 00) WYDAWIIG YI AN staptaagtagy tetatiant padtykadees adel td Frei pre ETRE eed el eed Qe dies gl es eyed bbe yt errere! wea at gd heed Eats Ete 4k ae gee ee AAR 3° 20 ° i] 40 ly c 50 N37C AWAY \AAAYAY ANuidds WNdshai\ds fy MWA Meade ekg ed tkathaadtd yatrt Ja. a kadt kt ; _ . ee HE HH eb eet a mire | +44 +H hy del Spee Hi Neel yi ea { Piet dat) bith See radth ee seeeqeertela ee be ad kath kk tht “AMAA = teh tea AVAN Nh NWN AWA ribs Wy vyw nn Sarlaaditaad§ ets gteted whee ye : Ok Zisuapeesert LI allt La debt Say iran TRL | HHL ane mek bs 4 we 4 q af att ay ‘ LEAK ak 3 d bee kh ddddhad 3 i = fiat | Mua “ aad J) ~N37D REF: fic | Fak er AMM AMSA \\ NTA 3) adadh Ke t4 deh adbd Us cb El Igy bgidkg +444] +444 +4 +4444 +H aeeL gate autasd agetat thet | gba tHe Lt Ltd [++ tirdy | put VI Hil | +44 +4444] 44+ tH J++] rill pi tb ol 441111 ky ty pit td thee ateh RQEeL 4h és Prd pay thak ete? dee eed cae dhg te eeee abet tek EE DATE: Ndi an yg Wl AWA) AYN aad “eeaed | akatad tidkad ad eet att aad raed 7 gad Ladd ++4 + 444444 FHEEEE FEFFEL HE) Fe. +b pettt + ++L+ pil +r t ft fl ditde4 rin rinel Fld yl ;4i4) \ |4 Vtly pag det bk barry k bE REE peeekh ake Eta ERLE bE REL wdtas by ced 10 .O nN o ” B ) ) J NOTE DATE: REF: 4 _f _” ee 10 VA sok lf on Ldkd WWM MMO | (3 aaddd kdbdd} aa tbattd “aid \\ SMM \\ Sin lade pe atad dtditd rdevhd 4+} I++4| Adit] Hi tbE wel + HEA} FTF} AFF H+ Fe gt read SI Zt both 3 rit fiyit aeli dl +444 rie AT Wiel baste bth ype ppp Ve bh L beek ake beedtl thet “tice radh td ett) tad dt thd DATE: S anny of ty perme By BR eel 37 yi a, Ve, Amer, Prk, Vi; (72). i“ l2 LsL. REF: 1 4 10 10 20 30 40 pam HCO) + inne 50|| aged st lI jm my Ceeser | Xero 4 pA A pe AA Ve 3| 7b SL ey NL REF 3 7A AHS Bae | Gyar| e | & | ce | & Vee | or Ie O° A-Vo | Al. 31/74 226 Sf 37A | 2AME E39 | B44 37B | 52/2 2. A-se<,| 21° | 35/7¢/ PE | io 32. | 20/65 a 374A | 29/218 rez) =| 36 378 | GlAzY ZH | A-P lel. [rere TIE az | 1h 2O. a 20, 37A et [aie (aes eS ‘| 37B@ | 32/224 KE any | 32a. | 136/24 -e2te gee 378 | ie/2ee | s7f Ver ees] 21 20 4 | T= Sd o42st| 20637 «/7/ yo - 20) 37 aa Aiew Sy -09509 | BPEL ASF (0S) 14S 37B 33/224 oT .2°425 ITO? u-P | 21 27/Id— -206 [usta 37BY) 27K) 374 : tea 239 (09/3 -%3Q3) «249 jase 2s 378 2Yray= VITO 06162-2634 -204 | Ve Y #0 374 sefye= Hot 29573-4987 3374 340 ‘42D 378 wog/zep =-HP2Q |-sl |. 7/¢7) « 47 fee -P | 21 27/1% =. 2b [2% 350| “259 32 7/63= °/43 0321 179.3 | © /Se2L a | || 374 wife = ZN — 0#33\ 2996 -22bb 207 1% S01 378 Mfr =. BZ -oS[4S7-2268) -/ RST ° Aee-VY, (374 eli/aee-B7QZ, WAG SSF] BSS io) 548 letra se HS" | [+e ze Hag 39 | 36] 348 a DATE: P-V, 37A 376 374 37B° 30 40 50 Ve DATE: fol (= REF: + rn | | Mae, UBL O! + [PCu7366) jobrep 2a, 3SE—-/ M tle + W370 oO BE-F Se P/(w4er1) O 3F-2) PME) | O 3 — ST WBC HW 2BEF O BF-3, FMB EFI) | e300 BREE ph@4 SK be~ 3-3 % hue. jofpr. SummR yoapite Welegertio prute — DB toc, lofey SPR See ST pte oD) jofze Ve Rae ah peered Det > Borhe SNR Kenic, } wil~s- § Reel Yom rons of Ar KA sg a “de ane Soe, ® 20 ~ Leek. oe te TLTWEIED |G /7/ ge SER SR a BOF TE RMB MATE. | 778 ote KV-6-0) The Russe, Ooh tT, we ee) pes Ugh oO T . awlis (Se . 30 ad bee brine Oealed ow Bike + WE s a ulie(+s, asa Ve. 40 50 N39 20 30 10 40 50 nO citer a DATE: lal? /sb W3236 UY a Blue REF: 1 2 3 4 6 | 77 8 9 10 Bodmpd- Vo aie.» V7 pPptto Jo l@tprt Wee Refract, 3 pete. 1 /2P/s6. if2za1s4. |S Fretted ow | B-o9 ba Che. ate due Beto _ pate, lo/als& REF: Oo M dee 2 3 4 5 6 7 8 9 W923 | w2¢0/ 3266 | 13 27) W3 265) 3236 w3iB3 | wr o tr |#tF|FF | LK W312 | oO rt | FF LF OTL KK +o-/ YIO fee | OM 4H Fr IKK | FF | Ww 224 3 ak RYT Ett ++) 8 tHe ise [ee [FR UT wrZAS od — Hr Lo WI2BF | ++ > t+) RP) KY + | w3237 (Te | oO Lee] RE) +t IRE wT tHE | CO LF KE | AR KE le M342 | xe | CL mE |) ERK eX 4%o~3. cers HE] OO) FA) el me MITTEE | OL RR i eK Ww F Sh) ft OD FAK | KR eK 2 2.2 « WR2HS ~opWr 24s pre Jorpahey ranerak Fe Legit - ; 50 _ ote. (2/56 REF: 1 2 3 o_o 5 PE Meek a yin 10 YZ) + Octane. Keel ated Deh d. B-O. fa V36- Paghe Raney ne ten Bi-o oe rz Joc, oto AS] Spy ten: —E- Avtdeanel egp nS 0 ia $- bm 2adogpe- bana pre- 5 ope pri, pRodre well. | o./erP force + 4nl- MaO 10 + O.W AQ. ONFG (324 /24 ml) Pett Ee outer | abe haat || w37236 +t + Mittz, O — ) lwrdeQ | ae a war0, O ae wie | CO | ~ wiito| O | & p20 wary] pe [| |wiaqso, 4+ | trey # = |lw3270) © _ Bc dd) 3234) + — | nlbo+ boast. —-) fwszy | & wits! o fe | Oren igi ¥ w3264 | © — w3237 CF | tn fre Bite x Be } | 3HB | a |e 1 | ¥7O > —— | solwit¢) | —+- oe | ees) + wit¢t7, CL 1 w3272, O i } tee) + lL | web Oo € Ww3267| +- . out: Rept wifts | + | } | w32e) 4 | t= py fed Wer irge 3 a0) w926F + | oUt aon Of & ew aliyy lac-/: X fi w3746) Ee a ane ( W319) 2 OM AGT, Ghee nn, OPE 8 Ke w3227 Coe -- | 2) ang seg tal A Be IS bie onpt Re w327!, CO i 3) of ne OWPGT be W726 TF 326 “we-f | W323F 42 ee "I J) 50) wiT4G — | maar ee Wo aa by udase wmaty¢4, Oo) | a Ort€ eK fo fot bees — At la3lsse.. Ww 31 3-q| WIZ 207 W326 Feretaabeel atl OMPet 10 40 50 REF: \ DATE: t/ /2 o/s G . ? 7 Nee AL er B~ AF 2Onro —-O, 10 DST ea + Nk Mr it | ww B229KX ww ‘\N Ww 3229Y4x WW |}w32244¥x wu wag k x f w3idole xX & W3leOye xX 4 Ws ws Ky A2ES —— 20f Ao | a 9 1 nad fe ; ede, C7 ne. prnnemy bho) ik Mis, ee Be ee tala ont pmegtt Mt Vo et Wik 75s hee N¢a-B DATE: REF: “Tere hue o_ doer Va Fp eet op Jon Anpeeka, [a HEMT PR Dorcas” Ur om M ne.) SA ke. tte al M Lec Q _— HER MTT Sad ng Ue meme RO oo wan Oph ake 76. | A An Ba emt maak L~ aarad Ante Tree Coe carlin tide ne M Ree. . Ve edutn, Moe? Product 7 C.0 Vel a te CT + |e Ue e+ | aL _—f— Y i Piet Ane E oe Ping By PoE apa be © . ~ pate: /¢ [xe [s% . REF: 2 1 3 4 Ss 6 7 8 9 10 A tei XRT) afl penta | Peto Kt AX yl) ole | unk 2) xX IA a-IAR., AHN 3, SG T,F) 7 DATE: NAX3-D REF: 10 20 30 40 50 Be 2-5 . ka. 50D TMC fa 20D Mire. TC TAC TAC —4- 1TMC Tae Tme | me J a — 33 | a) » ) sero ALP per coknning prrkekmphg - Stat B-Voge tt Tb, Dae | a an bh Loe mae ens hoe oat d- pe . Cree 3 Harsh on by due 3224 >/Y4-0 327 | we cD oS ss ™ So Oo Suk te pS ct Wels? week cate: 12/3 /5K ner: AALS 1 2 3 4 5 6 7 8 9 10 dee, (itz) TLIB fen ~ eek HEU be MN Toa Xo K waae7 Van) o4{nQ. pet pow ms Dm+ | dee. | Siale Ble. tol 49 | Ne Mea 322 ) 243 » 32279 =) LTY? Y| 20 49 46 v +47” Tory open aryey~p ~O~*dN ep 3r74¢O|)| 324D 30] 32357 3266) -—— AY Melied_ /T4S~ | ro Lager SB“ ¢E yy (q~2| Y aol /eya | 7) 3239) » — 3764 and 22497 20 be. | 323 so] 326 A aU ep Pegg 1). ode | - Pcp) AP eel SPR 9 frat tt pate: IX/t(S6 | REF: ; 2 3 4 5 6 7 9 10 Tian. 2k) <2 1% agen He Olek fn Ue + 0.f wh Fe ling Porm A phe ( cas antl /pete), 4PM. Ath +k Lf broth . Ep ene seat san Ba ie pe ~ls-~s-& |C~2H- : fe & W323h Ww3236F, LJ age} | SEO > a O Lt + S- a e 32297 Ss _— O * LY ES S a © __ 3/7¥¢L wn. Mae ts + abe rere Bderrtemn P46, Ss AW © . at— "47 < - . o 3144 | S ~- O.| * 329/ S A- Ee F240 S ow © KKe-3 | 315-9 S a © 3H Ss ne a 40/ 3237] Ss A a) (143 S . A— © T4.S Ve dapatg. wr © \ \ ee R-6 Por RR REF: 47-1 WX 10 47-2 | 20|| 30) Bu 2243- [Bash 34-2 | 393.1 | 3272 . | 3236 1440 r 327044 ls270b0 7 |327/ 32721 1349—) 1 A+ 37-2) | 31-2 | PM (2243 © (| 22MS7 aos.) 3234 | BLO BAD it s-o | ts 322] Site Daeyup BUDS 32239. r 32424yy 3/47 0 SI 321S7 sts-{ owe 3234/7 22ys-f + | Co e ols ef i it%/ (74S LTE i749 57 LG 1t¥g™ |Jaese tee It44 (7 bok, 32372 Lite] 3 224 Lolo L150 WS] Baby 3p (ie ZY Soe 366 ¥+ Areas Kad] MET ! DATE: REF: 10 : 7a) wy BeeaQ y | ‘Fi ‘k i++ t 4 SB [ ttt ap rtreetliite es # bp N i ri ltetatlarae tiers —“ QS PHt it htt titete \- rtt+ttt tt lttte z! tt+typttae i tet tye reg. as 37 iWdws” (tae At4G 3/46 | i9GG 3269 52.7 O 332 +t tet pst aco Sai Sie Pl hr] tt). - Tt FRSEY 100 0.00 DATE: VOf?2156 4 REF: VET A Gi Ade +Y 10 | + athbzM ae a oO Cy OQ + 410 1 3g¢eg 3 ‘| 39-lF XT oo t+ s YELIQQOUVRQ9Q t t 0Q0090 | : tae tT a4 gs 9 YQQ8O9 (3230) +t 0 0 | i “ot 37-3 326€ 3267 #266 32¥0 w+ 8 sae Le dee | pee, [ot War ee cenate. te tpl drop DATE: /2{t4 [sb REF: 2 3 7 Yea 7 W(2-2 || Fe /44[ 319 37 (14S ye 37 144957 (B/S wiz, & —— = tL 7 | 3/ j14 BeBe (fd? (a | 32 140° Ie) 137 — 16 |32 IT i ot en er [tol FO Let M6 (66.5 2M (Sb me Jer. \t Ts: ho pate: /2//3/5~L, i < . REF: V9 & \ J i 3 w326% | 2270 3 3/32 YIO _ w 3229 w322,| Ge |. D194 | fA” 174710 Br4¢Ga 7T46 Osid¢ 34-7 349 —zm0 39-3 “940 &) Laff. 1443 /t¢Y (947 3237 H3240 3266 3 2¢ S40 5) we 326 | W370 [W327] 9 “BN 24> 223° 3239 3t4F NYuo ; sere Bist als7 323° PK Tools coef ote oo 0% et Aoobes Og oce OF tT? ~ 0000 Gt9 sopsovaceH 3240 3266 3 26 . 32694 Hat th | | Sy S804 voit[eooot?o + T ene foe fo 6 . Cc eS = we: Fyo foo00FtoF = a Tank 3237 39-/ 3 9-2. 37-3 x wl hud o~ YOCQ roF HOLD & FTEPF TO +T w327V4 treet dh a wa272h eect Hain Ht Hort REF: T v5) y/oT (Ene. 8 oO w3\3>] Y lox” w SW Yio 1O a 10d” Sl tf WZ223 W 3120 w322}| WET) wle4s7 30 _ 40 50 | © OOK oH oR ly toe ws| — DATE: won (SL. ‘REF: Bhee | Mane M Bae | Toe2 4 4 5 6 7 8 ,2 10 Lo a OSE It eaten Gleabete| miyey OO Té CY¥)0) Myer oO- récyjo\| ee O° Pw) | 4 withE LO W323¢14 | de 397~) VE) 39: 2M6 - i + + QeVIO-V9IFFOO-GSOFO BE, wl6qlg | waeog wttss | | pace) | Pl ke | wI2oe wad | wI237] mete eee mou ° | Ss” l O & / © /O / ° Oo 3 ints” O Ue IT ~ 4 ve MS oe : Oe hye A , —s =! 6 oe pate: /L/1E /SG REF: _ ; 2 3 P —_ 7 8 ° 10 M fp Yios~ | = oe. | (2B | ep . SRF Let Of. KAjoQ.H,O. . Qelel (dwp FOXE ALe anh phdeM foc. JS . re 30 | ep ae | 3M wat! sD IBLF tse & 24 | Wt] | Zoo At AY em | STZ |. LS (Pl(waryolh) ~37E7 Shure?) (tei) 10 TRO |2é |waruih 0 (waswe) PtH) 290 |[ 4D 27 Ww 3087 2. (woop), 20 l) 99s TT tee oe w3134¢| 200) LL q 307 | 150) 40 f 3/46) (50) ST — 3/5R_, FO 5 oO a 3/56/50, GE I/O ie RIFT 2e0) Tl | J W3/53 27 [UAPteninc aonb) wr24t 6 | & | & w3IKT) 200 2&0) w3i4e| _ ee W334 | LOD | =e vo w308T TD | Ze = WE [Oo = — 3/52. ZO | a 3/56. TO | -T aa 3146F7 O 33 1390 _ Second report on lac- study N. Morton Jan. 10, 1957 New stocks. Five new lace stocks have been prepared by UV-induced mutation in W3236 and called W3267,8; N39-1,2,3. Vs stocks were derived from each of the Hfr lac- stocks by selection in broth, In future, a ve stock will be used as source of new lace mutants to assure identity of the ve marker, Lac~ stable derivatives of the two allelic mutable stocks W3120 and W1950 were obtained after UV treatment (N36). Of 10 non-papillate colonies of each type tested, 2 of each stock failed to recombine with other lac-1 mutants. These colonies were labeled W3269-72, %. Mapping of V6, lac, P, A cross was made between W1366 % lacy (VESTER, )"Fe and W3236 Hfr-1 P- M- on D-O+proline. The scoring of 218 recombinants gives as the most likely order i (Her) V6___—_idac P vt) (TLB) 35" 15 Be “31 38 In several other experinents. was | some selection for P+, but the same order and approximately the same distances are indicated, ONPG tests Single colony isolates from B-O were grown overnight on a rotator in YZ broth + 0,5% lactose +0.5% glycerol, These cultures were spun down, the pellet resuspended in 1 ml. water, and the cells autolysed by shafling with 1-2 drops of benzene. To 0.1 ml. of this mixture were added 2 ml. of an ONPG solution (30mg./l00m1,) and the color read by eye after 10 mimtes at 37° C, ALL of the single-step lac-1 mutants were ONPG+, Of the two-step lae-1 mutants, W3159 and Y70 were also +, but W3229 and W3269-72 were ONPG-. All of the remaining lac mutants were ONPG- except W3267, which contained reversions, W3268, which is a weak fermenter of several sugars, and W3239, a lace stock received from Borek, Lac-3, lac-5, and N39-1,2,3 were not tested. Aliquots of all stocks were tested for lac+ reversions before autolysis; only W1950 and W3267 contained a detectible number of reversions, Fermentation tests W3268, N39-1, and lac-3 ferment galactose, maltose, glucose, and arabinose weakly or not at all, N39-3 does not ferment maltose, but does ferment the other sugars, Lac-5 does not ferment galactose or maltose but does ferment glucose and arabinose, The remaining lac- mutants will ferment all the other sugars tested, Crossover suppression in two-step mutants Of the 2 ONPG + two-step lac-1 mutants, Y70 shows the “recombination pattern as ¥53, and W3159 covers all of the recognized lac-l region except W196, W3229 and W3272 cover all of the lac-l region and presumably extend beyond it, since they are ONPG-. However, they have not been shown to be allelic with any ONPG- single-step mutant, W3269-71 do not recombine with lace2 and lac-l; or with the majority of lac- mutants of unknown location, Either the region of crossover suppression is large » or most of the lace mutants are in the neighborhood of lacel. fransduction tests If an M lac plate is spread with two drops of an F- lac- protetroph and half of the plate respread with 1 drop of a Pl lysate obtained from a non- allelic lace Hfr ve by the Lennox modification of the Adams layer plate technique, then a yield of from 20 te 300 colonies will be obtained with Pl at a time when the control half of the plate is blank or has at most a few colonies, Allelic lysates give zero yields, In all combinations so far tried, the proportion of v, among lac+ colonies is less than 1%. No per- sistant heterogenotes have been obtained. | Recombination tests More than 20 crosses have been made of Hfr Ve M- lece X Fe lace prototroph 300 on M lac, the colonies being crossestreaked, without purification, against T6 on M lac, The Hfr parents have been W3229, W196, W3146, and W3120, In every case, the proportion of vs among the recombinants is less than 50%, with no evident reversal of ratios in reciprocal matings, Discussion So far attempts to map the lac-1 region have been unsuccessful, This may be due to multiple adjacent crossovers in pairing regions, to proximity of the Hfr-] elimination region to Ves or to heterogeneity in the location of V, mutants, It is not clear what the most expeditious way to investigate these possibilities will be. Tentatively it 1s proposed to use W3133 as a universal receptor for all Pl lysates and as a universal F- for all Hfr-1 stocks, excluding lac-1 lysates and stocks, This procedure is suggested by the fact that W3229 ve gave no V; recombinants with W3215, W3151, W315h, and W3112, indicating perhaps that there is a V6 locus closely adjacent to the crossover -suppression region of W3229 and W3133. If at least one cross gives an excess of Yes this will give some hope that the region can be mapped with existing stocks, If there is no such cross, the most hopeful possibility would seem to be that the Hfr-1 locus is responsible for the mapping difficulty, and either transduction with F- donors and receptors, another fertility system, or the use of P as an unselected marker may be tried, To introduce P or another Hfr into existing P+ stocks would be difficult, nes pate: //,0/- TZ REF: a ‘t Bop 3 HPs eed YE PEE OTE Q- Dane ~ phe nw bw tg oe C 4 * - - a. Of 49 | W323 Ln foe-t. HxOw!) a.48 ft WE. 4 10 bet M4 Og aK . i cdunnyo bo? oe 2 re fa dee — Cre ender 2. ABP Peete SN is 2 Ot me @ y Demat Ver | ver comm emp fT . weg. (opetiD) QD L370 (ho. Ab.) 2 was tl yw) GC). t [a w yy we V22BB CA LHIEO CO ©, PIV EA AYAD |. wT 7 pe . Ast 7 £7 ~ Nv WW o~0 _,anEeT iad pens f Ki o Oo~ nb > & > XX | () 4 N oO SQOGSebQseo rs 8 3 iF &S 00. |_| I] 3) @} 6 0 W223 6 (or = NED nthe _ | Ce c2 Leen oe 4 . dN TT pee hee eS a? ; . ; . - Len WS? We — xX W247 " aq) f MS 50 ~ : wrayer @ oo, = , oe Wasp Key fa. Weare an | 8 HOM YO? DATE: S0/S 7 REF: nes A 1 pod Hee Ve ty Saag Uy 2 1 (< ¢ t - ¢ ; é) “ , . f Oo» “f ° (3 ee atte 7 w32s7) OD ws 3/5 wala} oo 4 W127 he 2 Be yt NBiTIE Ws 22g $1 Ve. KEN: w 3237 F-A + N39 FO KE — x ng RS SY Bil A - ah > 7 ~ att t Ms ot iP i” ~ Ay AL - (i } Cc f ~ 1OOQ ¢ WBA BQAOLG & h —_ —s (Poo t “owe ae Ave ~yt Od / ra ry Lf, Sd ” onan Spey aD we ot | _ Ft + Fl 38 q] 30 Is #4 DATE: 1/23/57 W332) Kk a YT. de 50}. B6VE yh Oy XM OY, +A REF: 8 ae famed 23 VES Pls. 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