Oliver Smithies:[00:00:00] Book d, which ended April 13th, 1973, the last page between the covers had a long list of all the bacteria, and bacteriophages that I had been considering in these experiments. It’s quite an impressive list, one, two, three, four, five, six, seven, eight, nine, ten — more than 20 different strains of bacteria and bacteriophages which would require me to remember my bacterial genetics better than I do now in order to understand. So Book e, which begins on April 13th, [00:01:00] 1973, and runs through to about September 10th, continues this work in trying to look for branch DNA replication forks, and get branches of branches for electron microscope imaging of the idea that I had published in my paper on immunoglobulin variability.
So, the first page continues the work in testing to get something which was called “D152 lysogen.” Looking for [00:02:00] a bacterial strain carrying lambda bacteriophages or lysogen which could be induced under suitable conditions, and perhaps then cause to replicate partly.
And here, we are testing on Saturday, April 14th, on page 3, testing various things in relation to this idea. The results on Sunday are described, and my state of memory rather incomprehensible. The general conclusion on the next page, “Abandon this approach, get [Weisberg’s?] phages, and select E+ from them.”
Continuing this work and [00:03:00] how to isolate the DNA by [Zim’s?] method, on Saturday, April 14th, page 7.
And here, we get back to lambda dv on page 9, lambda dv from D, with a nice blue comment at the bottom, “Worked fine.”
And here, on the following page, I’m back to considering histones, and looking for a reagent for finding the nearest neighbor in histones, with a rather long insert tacked on, on considering histone pairing, [00:04:00] June 10th, or possible pairs, permutated, permuted, 1-2-3-4, 1-2-3-4, etc., all possible pairs, some ideas in relation to the arrangement of histones on DNAs that had been interesting me for some time, the possibility of a super-DNA memory, which of course we now know quite a lot more about, the modification of histone, and the passage of this epigenetic information between different generations. This idea was festering in my mind, but never quite reached a conclusion.
A fresh [00:05:00] start on the lambda experiments on July 17th, after the vacation, trying to get (inaudible), with a diagram of what I expected to have happen on the following page. [00:06:00]
Looking for suitable lysogen to liberate phage under my control, page 17, received from Bob Weisberg various mutants. [00:07:00]
Digression from this topic on Saturday, July 28th, page 21, test of diamidine ester cross-linkage of hemoglobin, with attempts to crosslink hemoglobin. Gel of the various hemoglobins made on the preceding page, on Saturday, July 28th, looking at the results.
So I would jump from topic to topic between days, back to the strains from RW [00:08:00] on Monday, July 30th, with a rather nice cross-linking agent looked at on Monday, July 30th, with a disulfide dimer of an amidine, as far as I say, amidine ester to use for cross-linking. And I went with a comment that I left earlier for Lois Kitze’s birthday. [00:09:00]
Continuing with these various bacterial strains on the following pages, and bacteriophages, test of RW phages, etc., July 31st, Tuesday, page 33, with the results of various strain tests on page 37. [00:10:00]
Continuing with the bacterial genetics work over the next while, and at the same time, retrying hemoglobin coupling, Thursday, October 2nd, with different cross-linking agents. The cross-linking with hemoglobin, and the use of the disulfide, as a controlled, [00:11:00] reducing and stabilizing agent, was, and still is a favorite topic.
Back to phages, etc., the following day, and so here, my notebook clearly showed that I was thinking about two things at the same time, the hemoglobin cross-linking and the bacteriophage experiments. Typical, on Friday August 3rd, phage cross complicated at this point, in my career, complicated [00:12:00] bacterial genetics. And, this alternation between the two things.
Page 66 is an example of what was going on. [00:13:00] Many comments in different squares about what was happening in the different crosses.
And the following page then, gels of hemoglobin, with August 7th, page 71, rough S-S interchange experiment, and now, this is then August 7th, 1973. So this is Tuesday, August 7th, page 71; we’re continuing. 1973, it just, [obviously?] goes back to my enjoyment of the disulfide bond cleavage and formation paper, [00:14:00] which I published in 1965. That’s one of my favorite papers as I’ve no doubt said many times. And you see then, the following pages, back to crossing, bacteria, and bacteriophages.
Checking problems on Thursday, August 9th, and the following page, cross-linking hemoglobin, August 9th, Thursday, and the previous page was actually still on the same day. So doing both at once. [00:15:00]
On Monday, August 13th, one experiment is cleaning up X-7734R, clean up bacteria work, and following page, Monday August 13th, same day, hemoglobin experiments with a crosslinker that contained six CH2 groups.
On the following page, in the same progression, both going on. [00:16:00] For example, on page 96, “The result suggests that after the disulfide interchange, the molecules are locked into alpha-beta, or alpha2-beta2 dimers, rather than alpha2-beta2 tetramers. Otherwise, hard to see how they could get faster.”
And next page, a massive diagram of the results of streaking various strains. Quite two-headed monster, as it were, two things at once. [Back to?] F-lambda plasmid, a lysogen, [00:17:00] F450(lambda), cloned from [name redacted], a good lambda lysogen. And different temperatures needed for growing the various strains on Friday, August 17th. [00:18:00] [00:19:00] Trying to isolate lambda phage DNA on Monday, September 3rd, Labour Day, using 450 lambda plasmid, with the result on the opposite page, and no DNA in the product on the electron microscope. Sperm DNA controls are good, so it doesn’t work. [00:20:00] And continuing as the book runs out.
Control lambda dv for D on Saturday, September 8th, to reestablish the methods are working, set up from an old lambda dv starter overnight at room temperature, etc. This is growing nicely by Sunday morning, will be fine by Monday. [00:21:00]
Bulk of 450 lambda and lambda dv, on Monday, September 10th. As we come to the end of the book, with the usual list on the covers of bacteria stocks and the genotypes that were available, again about 20 bacterial stocks there. That ends Book e.