Professor T. 1. Sonneborn
Indiana Univaraiw
;)epertaent of Zoology
Bloomington, Indiana

f ddqed anararing your letter for a few days to give aqyrscslf aome

                      X was merely seeking infor-

time to think it over since car%ain features of it pusaled me greatly.
X,et ae first state that I am indeed sorry if my lettar caused you my
conaern.
nation.
Amrican article that Lindsgrenian independent experiments had been per-
formed on the malibioae pl-Obl8rP and was naturally anxious  to find out
about them. I waa apparently in error. Hcnever, I. was not alone in
reading thia irpplication into your staterslent.  SemrsJ. people who had
not 81 yet taken much etock in the Lindegrcsn stateaaent have already aom-
~auniaatetd with me on the basla of your remark and suggested ratbsr glee-
;Fully that thie settled the iersue.  It my well do 80 in the mind8 of
laany who might have skill had SQW doubts, but I fear that I shall have
to retain some rtmervation~.  %is labst development is quite typical,
it 5eem to %e, of the whole mfffafr.

I did not intend the% it should.

3 perhapa wrongly drew the conclusion from your Saiantific

  You raised certain questions in your letter and I shall try to
answer thee as streiqhtfomardly and honestly 89 I can.  If, in doing
so, I wpear overly harsh, please forgive me. Put it down .t;o the in-
teneity of my efforts to qet at, the truth as I see it, with prhap
~ome attendant sacrifice af the frills and nicetfes of polite inter-
course, 1 am sure, Tracy, ;you would have it no other may. In the
Brocess, f shall probably raise questions of my own to which perhapa
you can help provide answers.
to entertain as reasonable have only social implications and have lit-
tle to do with the scientiific amits of the matter and hence f am re-
luctant to accept there.

'he only solutions that I have been able

You start your letter by the quotation fmm L. and L., C. 3. W,

Syplposiun, Vol. 11, as many other peopls tislve in recent revicrsra. me
quotation is certainly correct although I ahould venture one modifica-
tion, namely, instead of reeding ftwfth the help of", it should be "under
the, obervation oft' since Michael had nothing physically to do with the
eexpritacant,
do cremes and microdissections.

By their very nature he couldn't have, involving as they

Michael's estimate of these experimcshts

uan be objectively gauged by the fact that he was unwilling to have
them published with his name attached.
issue.  What has surprised me is the fact that for over a year and B
half now, the actual data of thorse experimcants has been available in
printed form and not one person to my knowledge, who has announced his
unqualified faith in these experbents has seen fit to examine the
actual results upon which his conclusions f.w should I say faith) are
based,  I refer to the data published in Lindegren'a book,  'Ifre Yeast
Cell.  NOW I know, Tracy, that, it probably is asking a lot of any per-
son to demand that he go through this mess in order to get at the faata
of the situation.  Nevertheless, it rould seem ta Be that the usual
rules of scholarly cam and competence demand consideration of any
existent material pertinent to arriving at an adequrate estimate of the
question being 1Usoussed. Particularly is this true when the material
in question is the only data available and it refers to an issue a9
important 88 the, coxu-eference to the aelibiose experiment would
appear to make it.

But actually this is beside the

1 must confess that I find it a bit surprising that people who

would not believe Lindegren if he  told them the time of day neverthe-
less accept without qualifications a reek of his on an important
isme, unsuppirted by any experimenial evidence.

   In any case, let us take a look at the actual data which doe8
exist on the question,
in its conception.
ixchnioal pitfalls.
time.
tion there are many thing8 we WQuld like to know.
nature of the strains employed.
in the best run laboratories dealing with thousands of strains and
hence cannot be accepted as identification wit,hout question.  One might
hope to gain some information QII the question by a careful comparison
of the reported behaviour of a given (i.e., given labeled)  strain under
standard conditions.
arouses one's suspicion as to their capacity to keep their strains
straight or maintain Ulea in a condition in which they reproduce the
remlts of a particular experfnent from one period to another,
am my cases which can be cited but let me mention for an example
ones which is directly oertinent to the melibiose problem,  since it was
the fir,t Doudoroff-observed experiment.
ter 26.  "When he (this is Mike Doudoroff) arrived, we were analysing
the matines of certain strains.
from the rather regular pedigrees shown in Table 26-3 we predicted to
him that only regular segrega3ions would occur because we had finally
weleaned upn the cytoplasm.  %e were thoroughly surprised to find the
results shown in Table 26-5."
denly ahsmging properties In the Lindegren lab from one week to the
next.

The melibiose experiment was certainly sbple

Ita execution on the other hand is fraught with

Some of these I did not foresaee or realfie at the

                    One of these is the

In estimating the comparabili$y or even relevance ,,f tc repeti-

                 Ubels or names can be mitched even

Even a uasual reading of the Lindegren book

mere

     I quote from page 10 of chap-

Since all these haplophasas had arisen

Apparently strains have a habit of sud-

-3-

Wow let us turn to the actual melibiose experiment itself which
                     It is these experiments of course

  I have discussed these very experiments in a footnote to a

appears on page 19 of Chapter 26.
which form the basia of the doubts raised by Lindegren and reechoed by
others.
forthcoming paper in the Proceedings of thar-National Academy of Science,
I as sending you a copy of thia paper, but A should like to note a feu
things about it here.  &e cross which was originally employed in the
mlibiose erxperiment was one which was characterilasd by the fact that
it invariably gave regular segregations of 2+r2- in all tetrads which
uere completely analpttd.  I should further like to point out #at this
regularity did not stela merely from the "a$libiose" pper itself.  This
cross was chosen precisely because it had exhibited complete regularity
in *e course of our meviaus investigation, published in P.N.A.S.,
Vol. 30, page 346.  This was the paper in which we first proposed a
mu1t;iple gene possibility for the inheritance of a ntelibiase sph.

?he stock used in the "repetition" does bear the number LSC, which,

except for a k,does not differ from that used in the tno papers to
which I refer,  But look art the unexpectedly different behavior under
norrnal conditions of crosgling and segregation 1  This cros~, instead of
giving the usual 2+t2- segregations which it always did in the past,
yielded only one normal 2+:2- out of nins tetrads analyeed.  Five of
the asci yielded a 1+$3- ratio and two asci did not poshless the posi-
tive allele in any of the four spores of the hybrid!  me Lindegrens
have by now apparently bacoae inured to such Nsurpri8eef* since no com-
mentsr are made about this result.  I should like to note rJomsthing
nhich you can emily verify by looking at the pedigrees we published
on pages 349 and 350 of Vol. 30 of P.N.A.S.  In none of our exadnation
of the melibiose character had we ever encountermhis sikuation;
excess positives, yes, a8 ha8 also Winge3 but excess negatives,  never.
In fact, as far as I know, this type of complete "depletion" mutatton
only mcurs in Lindegrenfs laboratory. Under the circumtances, with
these data available, 1 do not think it unreasonable to entertain the
suppoaition that the strains used by Lin9egren in the "repetition"  is
not the one derived Proa the primary analysis of the melibiose pedigree
and which was used in the subaequent investieation of substrate effects
on aegregation ratios. Or do you think mer unduly hsrah in th? criteria
1:have set up for contparability?
the type of crom to use in an experiment designed to examine the effect
of substrate on "Mendelian" segregation ratios.
ing the cards just a wae bit if the strain can't even keep its genes
s traightt

In addition, I hardly think this is

It is certainly stack-

  We have not yet exhausted the rewdirkable! features of this so-called
experimont.  If nuw we look at the "experimntal" cross in which meli-
biose watt maintained throughout the cycle, we find the amasing fact that
an entirely different pair of spores is employed.  Inistead of LSC one
finds LsB and inswad of 20, 25.  And 89 a matter of fact, 20 is not
even a sister spore of 25 which wtil9 used in the control cross in the
absence of melibiose.  'his is a rather queer way to try a repeat ex-
periment of the effect of subbstra- on the transmission of enzymatic

J

capaciw through aeiosis.
whether the experimental is ooqwable to the "control" cross.

At #e very least it raises serious doubts

   %en, if we go still further and exaPiine the results of the CrOSS,
which was done in the presence of sub~tsa~, *FB find amsingly enough
that out of four coaplete tetrads analysed, all four gave 2+K2- inaJtaad
of the one out of nine obtained in the "control" CFOSB. Xf therefore
we do accept the experimental crosa as conparable to the control, re
must enbrain the p0SSibility that the presence of melibiose hss had
an effect on *&e segregation of positives since the probability that
thir uo Id he due to chance rauld be smmhem in the neighborhood of

1in1. d:

Actually of course, I don't place any felt. whatsoeter in thoas

eqrisnents in vim of the ray they were carried out.
how they can be seFiously entertained as constituting data with which
the earlier results can be corapared.
exptsriwnb with subsbatea other than melibiose described in the, same
section as 1 have had no exprience with thean.  So much for the Panow
rep ti tion.

I do not see

I em make no comment on the

  At the? end of your first paragraph you point out that you "need
not cite far you Piingels papers which certainly cannot be ignored in
this conniaction, even if you criticize theia."
confesrs you have lost me.
accept Lindegren t s conclusion #at the melibiose experiment does not
exist in fact.  %en you inmediately cite a paper which suggests  that
the facts are valid and indeed offers mother explanation for them.
don't quite see how one can propose m alternative explanation anti at
the same time deny the validity of the faa& which are presumably belng

interpreted. Z probably am iaicsaring sclmetating hem. Other people have
used this combin&%.on an %e.

At this point f must

In the first part of the same paragraph you

I

I still don't get it.

   Agaid, I apy be mong, but in view of my analysis of the Lindegren
"repetitiontt and Wingets findings it appsars to me highly probable that
we did discovler in 1945 that prior contact with substrate Influencd3s
the observed seqregation of phenotype and that this fact was oonfimd
by Tinge and Roberts in 1948.  Certainly Tinge is of this opinion as
he explicitly and in no uncertain terms states on three separate scca-
siom in the Finge-Roberts papers; once on page 3x0 in the fourth parr-
qreph lnnd again on the same pap fn the 1-t paragraph and finally once
&ore in the fu% paragraph on page 313.

After spending trco years investigating his phenoaenon I am ahat

certain he is correot and that we were dealing in the nelibioae experi-
ment with the same phenomenon.  ?y eorreswndenca with %8inge describing
our reaulta &iring the course of the exprimnts confirms me in thie
o~inion.  f txfght add that Wiinge ia beginning to wonder about cytoplas-
mic factors in connection with ensymatic adaptation.

-5-

   In conceiving #e melibiose expriutents, I did not foresee the pa-
sibility of such in phenomenon as long-tsnn adaptation and the experitaents
were not designed to take care of this possibility despite, the fact that
we held the tubes for a long period of time,
past few years with this phenomenon,  this is not the method. to employ
for detecting its euistenceb,

From my exprience in the

  tplinge an? Roberts made an extremely significant contribution but
not, in aly opinion, for reasons they and many others suppose. In any
case as you undoubtedly have clearly seen %e appearance of the 'Ji'inge-
Roberta paper ralmd arerioucs isawaa with the concluaiorls drawn from the
melibiose csxperimenta and indeed raises doubts whether on experiment
along these lines can be designed to test for the erxistence of gene-
initiated but independently self-duplicating entities.
however auggest how this may hs done.

Their results

   In my case, I decided to devote my major efforts to an analysis
of their phenoroenon for I felt imtll the underlying basis of this was
rerally clearly mderst~od there was little hope of constructing dsfin-
itive experivents.  It was not, an wisy investigation and it took us
almost two years of preliminary work to obtain the condition8 which
permitted an adequate analysis of the obvious pxstbilities.
time urn not completely rrasted for in the process we acquired the ex-
perinentizl "feel" for the naterial and phenomenon which is indispen-
sable in biological work,  It has pnt 11s in the pmition of being able
to ask and -mer questions about gene action and enayne foma+A,on
which we either did not think of or could not hope to solve prior to
this work.

This

  I have detailed these efforts a bit because in part they anmer
a qwati.on raised in the third paragraph of your letter on why I al-
lmed four years to slqxw without sayiug ctnfl,hing about the melibiose
exmrimen+-,

In the first place f naa bwy join% %ho only thing I thought was

pertinent to the issue, nalcnely gathering som relevant facts.  Perhaps
3 was naive about this but I felt that expressing my opinion about the
adequacy of the Lindegren repti%ian would be rather pdtntloss, despite
the fact that I had good reason3 to be suspicious.  Neither 3 nm any-
body else was in a position to estimate the validity of Lindegrenrs
remarks of 1946 until the dater upon whiah thsy were based became avail-
able and this took three years.  In the meantime Wingets paper appeared
in 1948 which certainly should have raised some? doubfx as to whether
L.rndegren knew what he was talking about.
tion was resoundingly settled by the appearance of his book.
prove concluaively that they haven* t the foggiest, notion of what an
experiment is and are completely inccmpetent to carry one through with-
otit the closest sort of supervision.

In my mind this last ques-

hey

-6-

As you well know, Pacy, I was hardly in a position in 1947 and

19h8 to accede to the deanand of nay public" to set up a micro-dissection
lab in order to repeat the mlibiose experiment.
conditions which exist here it has taken me a pear to get such a lab
built and equipped,  It will take another half-year for us to become
sufficiently proficient in the, routine technique8 to begin to put out
adequate data.

Even under the ideal

And now I should like to consider the questions you p988d in pur
a) my didn't I write you when the Heredity article appeared.
b) Thy didnt t I write you after seeing your course notes, which

postscript.

by the way are tremendous.  1 was delighted many times with
the penetrating nature of your analysis of various problew
including I might add the, yeaat work,

c) Why didn't I talk to you at Columbus?

Hfth regard to (a), please understad if the nard nthem" had not

appeared in the Scientific American article I would not have written
you about it.  %is term with its implications did not appear in the
FfereAity article.  Hence in the latter you were expressing an opinion
with no added information.
yotm or arqbody else'@ opinion about an opinion.
surprised that in the Heredity article you did not consicier tho data
existent in Lindegrants book.
and pbblishing I conceived it possible that it was not yet available
to you at tha time af writing.

I do riot feel that I can profitably question

I mst confess 1 was

But what with time lag betmen writing

3i'ith regard to (5); this I find the most surprising of all.

Except for details I 6m in crmplete agreeaent with your discussion of
+he data you consider on yeaat in your notes,  I don't have the notes
available now since I returned them sow time ago, but a8 I recall,
you very correctly pointed out *hat In view of Vfrigera results and
interprn+ationa it was difficalt to understand why Lindegren could not
repeat the melibiose experiuent.
experimnta had been properly perforraad, whether they had been adapted
sufficientLy.  Betmeen the time of writing these notes and your review
articles you have apparently aatisfied yourself that these reservations,
and tfie apparent contradictions hetween Fringe and Robarts and Lindegren
no longer existed or were not crucial to fh~ discussion.

You raised QU~S~~CXN of whe"&er +As

  With rogard to (c) , rightly or wrongly I got the rather strong
impreasiort. from our brief cmtacts at Colurdbua that you were preoccupied
with nany things at Coluhue,
one of us could have profited Sron a discussion in the a+hoos?hore that
prevailed there.  The few frequently interrupted exchanges we did have
did not encourage me to entertain any other vim.
reason that I suggested that X pay you a visit in which we cmld go
over our researches in a more relaxed and rational environment.

I felt it highly unlikely that either

It was for that

-7-

Well, I must bring this letter +& a close.

Before I do so there

are several remarks I would hike to &e.  It my indeed be probable
that in the 78 or so papers that I haw, published mistakes have been
-de. 1 do not fear the committing of BOBS 9-1 the future. I ham
never omriidered that the proper functions of any erember of society,
and least of all a scientist, should be the jealous guarding af his
reputatfon to the and that when he dies It may be said or hiHi "he never
had to retract either a theory or 8 fact."  Piis kind of. epitaph can
be mitten for a multitude -de nu% by virtue of lack of ability or
otherwise, bgt can apply to relatively fenr who, by their creative
efforts, have fruitfully enlarged our knowledge and imderstanding of
nature. 19 I make an error 1 will admit it. Hosever, I will not let
fear of cornwitting one paralyse me into inactiviw.
dduslngd if I will seek aalvaUon from the! Gods of genetics (or my
others) by cmfssaing sins they presume I have committed.

But I will be

This has been a painf'ul letter to write and took mors time and

effort than even its Snordinats leng"$l wauld indicate.
ne over ground I have traversed my tima fruitlessly in the past
four years.  I am always le3 back to the rather bit%r conclusion that
it would have been better for the field had I never conceived and per-
formed an oxperf3en.t I had looked upm with sorno pride.  It has engen-
dered only irrationality and confusion and all far* heartbreakingly
irrelevant masons of incowctencs.

It ha8 taken

I how my efforts xi11 be or' some help to you in arrivirig at; an

underatanding of nty views.
I do not expect to pmauade you to my viewpoint by this.
as these tire not solved by such letters or semi-popular articles or
t~correctlm~ le%ters to the edi+&rs, Zfiose aspects worthy of our
attation will be resolved only by #e accumulation of more informtion
and the design of tha propr experimnts.  %is i am trying to do  b
the be8t of ny ability as I knarr you are.  Our job is not made easier
by the backpo?md noise.

If not, they will have been doubly in vain.

Issues such

Let me end by saying this, Tracy, you are one of the very few anen

in American Biology of whom I think highly eno-qh to write all I have
included in this let+Ar.

Depraseedly yours

s. Sgiegelnan