Let's get started.
So I'm going to finish up energy
today.  And then we're going to
begin sort of section more or less
we'll call it molecular biology but
it's sort of dealing with the issues
that revolve around the discovery
that DNA was the genetic material
and then working through how people
understood how information got from
the DNA into everything else,
how things were regulated.
There are an incredibly large number
of important discoveries that form
the foundation of how we think about
biology that are going to come out
in this next section,
but before I do that I just want to
finish up this section that I talked
to you about, about energy.
And I hope as we go along here
you're going to see how some of
these sort of disparate parts of the
course begin to come together.
Almost everything we're going to be
talking about now is going to be
needing energy such as replicating
DNA or making proteins and
all sorts of things.
And those are driven ultimately by
ATP.  And what I've been trying to
talk to about for the last lecture
or two is how the cell gets the ATP,
the energy money that it needs to
make things.  We talked through
glycolysis, this ancient,
ancient way of getting a couple of
ATPs out of a molecule of sugar so
well-imbedded in our genetic makeup
it's in almost all organisms.
And then I talked last lecture about
this other principle which must have
come up very, very early in
evolution.  Again,
it's used by all organisms.
And that's the principle of
capturing the energy that's inherent
in a proton gradient across a
membrane.  And I talked to you then
about the idea then the way it
worked was that the cell would have
something in its membrane that would
be a proton pump and it would pump
the proton from one side of the
membrane to the other.
So it's working against the gradient.
So it's doing energy.
So there are a couple of different
ways energy needs to be provided.
It could be provided by some kind
of light energy,
and that's what drives
photosynthesis.
I'll say a few words about that.
Or in the case of respiration with
the oxidative phosphorylation,
I showed you how it was the
electrons sort of descend in
stepwise fashion down from
one state to another.
There's energy given off,
free energy is available, and that
can be used to power the pump.
And once the proton gradient is
made then the cells can turn it
around and use the energy that's in
that proton gradient to make ATP.
So in respiration remember the trick
was then to take those two pyruvates,
burn them all the way down to carbon
dioxide and water,
make as many ATPs and NADHs as you
could, then take the NADHs,
use them to make a proton gradient
and eventually,
if you will, convert everything into
ATP so you've got it as energy.
Our cells do it.  That part,
respiration and the oxidative
phosphorylation is done in
mitochondria, which I said sort of
came from bacteria that were
captured at some point.
Here's a picture of a mitochondrion.
It still looks more or less like a
bacterium.  And I showed you the
little parts that are in there.
It was funny.  Right after lecture
I went back to my lab.
I picked up a recent issue of
Science.  And I opened up to a page
that said something about rats that
had been bred to be very poor at
aerobic exercise.
And it went on to talk about the
health problems they had.
And there was a sentence in there
that they think the underlying cause
is by breeding these rats and
selection four rats that are poor at
doing aerobic exercise.
What they think it all stems from
is having very inefficient
mitochondria that don't work nearly
as well.  And that would make a lot
of sense.  I was going to scan that
article but we had some technical
issues this morning.
Maybe I can show it to you next
lecture.  OK.
So the one last thing then that I
want to do is I want to say a few
words about photosynthesis because
that actually preceded respiration.
Respiration couldn't evolve until
there was oxygen in the atmosphere.
So probably the first use or
certainly one of the first uses of
the proton gradient happened in this
scale of evolution and put here
somewhere maybe 3.
billion years ago or so when what I
had called photosynthesis --
-- release I on day one in the sort
of trivial fashion.  This
is known as cyclic --
-- photo phosphorylation.
And the principle is relatively
simple.  It's to capture the energy
in sunlight --
-- to make a proton gradient.
And that then can be used, as you
now know, to make ATP.
So in order to capture energy from
sunlight nature had to evolve
molecules that are able to observe
in the appropriate wavelength
range.
You know the names of those
molecules.  Chlorophyll,
the come in two principle species.
You don't have to remember the
structure.  What you can see is a
lot of conjugated double-bonds.
That's how you sort of tune the
absorption of a molecule.
If you want to make it absorb a
longer and longer wavelength you
start hooking together double bonds.
And you can set it up sort of you
can get a molecule to absorb at just
about any maximum absorption,
any wavelength you want.  So
chlorophyll is able to
absorb this energy.
And the principle of this is what
happens.  You have this chlorophyll
and it absorbs a photon.
And an electron gets excited so it
basically moves to another orbital.
It's farther away from the nucleus.
It's easier now for that electron
to get lost than it was before.
Now, if nothing was happening that
electron would eventually just fall
down to its ground state and you'd
lose all the energy as heat.
So what happens in photosynthesis,
though, is that the electron falls
back down to the ground state again
in a series of steps.
And how this happens,
electrons are basically getting
passed from one carrier to another.
And the same principle as we saw in
respiration applies in that at each
phase in here a proton is pumped.
Now, the out and the in are reversed
from what it says in respiration.
You have notes on respiration, or
should anyway.
But the in and the out,
as you'll see, it's sort of an
arbitrary.  You sort of take a frame
of reference and then something's in
and something's out.
The point is that in both of them
protons go from one side of the
membrane to the other and you get
more on one side,
you pump them in one direction and
they flow back in the other.
And the in and the out is sort of
an arbitrary way of describing
what's happening,
but in both cases the key thing is
that you're pumping electrons out.
And then these can be used to make
ATP, as we talked about with ATP
synthesis.  In this case the
electrons eventually end up back on
the chlorophyll.
And so that's why it's called cyclic
phosphorylation.
What you get out of this,
as you can see, is ATP.  So this was
probably a really big deal in
evolution because the current
thinking is perhaps there was an RNA
world that's still sort of being
debated.  At some point it's clear
that somewhere around 3.
billion years or so something that
looks sort of like a present-day
bacterium arose,
probably eight molecules that had
already been made in the sort of
primordial soup,
but when those started to run out
then it needed other ways
of making energy.
It needed other ways of making
carbon.  Here's a way of getting
energy, but what's available to make
more organic molecules is only
carbon dioxide.
And if you remember that little
thing I showed you of if we're going
from a methyl to a hydroxyl to an
aldehyde to an acid --
-- to CO2, that direction is
oxidation and that direction is
reduction.  So if we go in that
direction and we end up generating
NADH because we're taking electrons
and giving them to something else,
if we want to go the other way if
we're starting with C02 what we need
to do is we need to have a supply of
reducing power so we can take the
C02 and get it down to all the
less-oxidized states that are
necessary for building all the
molecules that we've
been talking about.
So making ATP was a great idea,
but the cell still needed to have
some form of reducing agent.
And what they used was they used
hydrogen sulfide.
This is at least one of the major
ways that it was done.
And so there is a very slight twist
here.  This is NADP.
It's the same molecule as NAD
except there's an extra
phosphorylation.
And the one with the phosphate on it
tends to be used in biosynthetic
reactions, but otherwise it's
exactly the same thing.
It's an electron banking thing.
And what this gave was NADPH plus
sulfur plus a hydrogen.
So sulfur is a waste product.
Here's the reducing power.  Here's
the ATP.  That's what those
organisms need to be able to
synthesize new organic material
without having to have
pre-made molecules.
A really big deal in evolution.
And the idea for making ATP is
based on this use of establishing a
proton gradient,
the same principle we've seen again.
Now, there's another possible
source of reducing power,
and that would be to use water as
the source of the reducing power.
But in order to do that you've got
to put more energy into it.
And this system wasn't able to
handle it.
But that happened soon enough with
the development of what I called on
the first day photosynthesis release
II, which is technically known as
noncyclic photophosphorylation.
Again, it uses the energy of
sunlight.  But the twist this time,
it not only makes ATP, it also makes
NADPH, it makes reducing power at
the same time.
So you can see that is a really
major advance.
If you can use sunlight to make
both of them now you're really
efficient.  So this is how this one
works.  It's related
to the other one.
And the first part is more or less
the same idea.
A photon is absorbed by a molecule
of chlorophyll.
It kicks the chlorophyll up to an
activated state where the electrons
are at a higher orbital far away.
It wants to come back down.  Energy
is going to be released.
So electrons gets passed,
protons get pumped from one side of
a membrane to another.
Except this time,
instead of coming back this lands in
a different chlorophyll that has
just recently lost a pair of
electrons.  But there's a new energy
input here that kicks this
chlorophyll up to an even higher
energy state than this one.
And as these electrons start to
come down the energy hill there's
enough energy here to take a
molecule of NADP+ plus a hydrogen
ion and give NADPH.
There's one thing that this isn't
going to work like a cycle or a
machine yet.  Anybody see what
hasn't been taken care of yet?
Say again.  Send the electrons back
to this chlorophyll, exactly.
However, the way the energetics are
structured now the cells were able
to take reducing power from here and
generate 2H+ plus a half of an
oxygen molecule.
And this would really be two waters
giving four hydrogens and one oxygen
molecule.  So what you can see here
now, there are a couple of really
important things about this.
It needs more energy.
It makes ATP and NADPH which leaves
the cell able to carry out
biosynthesis.  And the third thing,
which is an incredible influence on
our planet, it started to generate
oxygen as a waste product.
And it's really a mixed blessing.
I mean oxygen is very reactive.
It damages our DNA.
It damages our proteins.
We have an amazing number of
defenses against oxygen.
But, on the other hand, as it
accumulated in the atmosphere and
organisms slowly over evolutionary
time learned to deal with it,
it then set us up for the
possibility of respiration which,
as you can see, is 18 times more
efficient than in that ancient way
of using glycolysis to make energy
out of sugars.
So that's more or less the story.
This part is called photosystem II.
This assembly of stuff is
photosystem I.
And I just wanted to show you this
next slide because chlorophyll isn't
just floating around like this.
As you might guess, it's bound into
proteins and things.
And someone has figured out the
structure of photosystem I.
It consists of 12 proteins,
96 chlorophylls and about 30 other
molecules.
And what it really does is it
functions as an antenna.
Some of the other molecules can
absorb it at wavelengths that are
different from chlorophyll.
And all the energy gets funneled
into the chlorophyll and into this
process.  And you'll probably
recognize by now that proteins here
we're seeing alpha helices and beta
sheets in here as part of this
structure.  So the first organisms
that learned how to do this were
organisms we now know
as cyanobacteria.
They're a kind of bacteria that has
two membranes like E.
coli and like the other ones that
we've talked about.
You're familiar with these.
There's the green scum you see on
ponds.  Here's a close-up.
Sometimes they grow as filaments,
the cells in a chain.  You notice
they're green.
They're making chlorophyll.
And what happened in plants was
that apparently something probably
related to the present-day
cyanobacteria got trapped inside
some early progenitor of what we now
know as plants and green algae.
And this trapped bacterium became a
chloroplast.  And it had all the
machinery necessary to carry out
this noncyclic photophosphorylation.
The structure of these things,
there's an outer membrane.
Just similar to what I told you for
the mitochondria.
There's an inner membrane.
And what's special about the
mitochondrion then,
there's another membrane inside
that's known as the thylocoid.
And that's where all the chlorophyll
is.  And the reason the out and the
in is a little bit confusing in here
is this part, which is probably the
cytoplasm of the old bacterium,
is pumped from what's known as the
stroma of a chloroplast which is
equivalent to the cytoplasm of the
original bacteria into the lumen.
So the chlorophyll that's in this
membrane absorbs the light,
pumps protons into the lumen
building up a proton gradient,
and then they flow back out in the
other direction and make ATP.
Here's a picture of a chloroplast
once again.  It looks an awful lot
like the bacterium still that got
captured.  All this stuff on the
inside, those are the thylocoid
membranes that carry out this
specialized stuff.  So
there you have it.
That's how cells,
the major ways that life has figured
out how to make energy.
When Penny Chisholm starts to talk
to you she'll talk to you about how
organisms adapt to various niches,
things that live in the bottom of
the ocean, things that live in
various places.
They all have to make energy.
Well, they all use some variation
on these principles I've talked to
you about.  And she'll then show you
how they're very cleaver at
extracting energy out of all sorts
of things by applying these
principles in different ways.
OK.  So what we're going to start
doing now is we're going to start
talking about DNA.
This is certainly a molecule that's
fascinated me all my life.
You should know from the first part
that it's built up of units known as
nucleotides that have a sugar.
It's a ribose sugar that's missing
one hydroxyl so it's a deoxyribose.
The sugars are numbered 1, 2, 3, 4,
5.  I showed you that.  They'll be a
phosphate.
And then one of these nucleic acid
bases, either a pyrimidine or a
purine.  And in DNA you find the
pyrimidine bases are cytosine and
thiamine.  And in DNA the purine
bases are adenine and guanine.
And then these subunits are
polymerized together.
In essence, splitting out water to
give you a polymer.
And I didn't emphasize this too
strong the first time
I showed it to you.
It's going to become a very big deal
over the next few lectures as we
begin to consider how nature had to
figure out how to replicate DNA and
all sorts of implications to go
along with this,
but there's a polarity to a strand
of DNA.  This is what's called the 5
prime sugar.  The primes indicate
the numbers referring to the sugar,
and the ones without primes are
referring to numbers of atoms that
make up part of the nucleic
acid base.
So if we're looking at a chain,
this is a 5 prime carbon of the
sugar, that's the 3 prime.
And so what you can see, this bond
which is really a phosphodiester
bond, the phosphate group has formed
an ester with this hydroxyl and with
the hydroxyl that used to be here.
So it's a phosphodiester bond and
it's a 5 prime,
3 prime bond.  It joins the 5 prime
carbon to the 3 prime
carbon up here.
So that means if you're looking at a
chain of DNA, if you come down this
way you're coming in the 5 to 3
prime direction.
If we come up the other way we're
coming from the 3 prime end heading
towards the 5 prime end.
So you'll see me saying 5 prime,
3 prime.  Now, as I told you, the
principle force that holds the
strands of the DNA together are
hydrogen bonds,
three of them between a G and a C
and two of them between
an A and a T.
And then they are a pair of strands.
And they're actually running in
opposite polarities.
This is something to contend with
when we think about replication.
5 prime to 3 prime in one direction
and 5 prime to 3 prime going in the
opposite way here.
And then, as you all know,
it's called the double helix.
So it's actually not flat like this
in space.  It's in a 3-dimensional
twisted into a double helix and the
base pairs are held together by
hydrogen bonds between the bases on
the opposite strands down the middle
of the molecule.
And I like this little movie I
showed you because you can see it
pretty well.  The nitrogens are blue.
It's easy to see the bases.
And the hydrogen bonds are right in
the middle.  There's another force I
didn't mention and it doesn't matter
for this course,
but when the bases sort of stack on
top of each other there's actually a
kind of extra stabilization that
comes from that.
It's a gorgeous molecule.
You all know it encodes the genetic
information.
We're going to be talking about it a
lot, but first thing,
you know, I could just tell you it's
the genetic information.
But one of the really big
discoveries in biology was that DNA
is the genetic information.
And a point I'm trying to help you
learn here, you know,
I'm trying to teach you more than
just facts.  And I hope some of you
at least will catch that.
I'm trying to show you how biology
is done.
As an experimental scientist you
don't sit down usually and figure it
out.  Instead you start doing
experiments and you get all kinds of
unexpected discoveries.
And, in general, as people work in
these unexpected discoveries
ultimately they come to these grand
new insights that,
you know, sometimes would have been
very hard to forget.
So the real question that people
wondered for a long time,
and we'll talk more about the
history of genetics,
but people knew we clearly had
inheritable traits.
You could see it in your kids.
People had been breeding plants and
all sorts of things.
Breeding domestic animals.
They sort of understood the
principle of inheritance.
When I tell you about Mendel we'll
begin to see how his thinking led to
the idea that the inheritance wasn't
just sort of like a liquid where
everything mixed together.
It came in units or particles which
we know of as genes.
And so the idea that genes had been
accepted certainly by the beginning
of this century anyway,
but nobody knew what they were made
of.  They were made of --
The major properties that they had
was they clearly encoded information
in some way.
They must replicate because one cell
could give two and on and on and on.
So if you were going to pass it
down in an inherited way they have
to be replicated.
And the third thing was that people
knew somehow they could mutate or
the information content that they
encoded could be changed.
Again, you could see that,
that you'd get something, an altered
characteristic,
and then it would be propagated down
through that line.
That was the principle of breeding
that people had done for ages.
And so they understood that.  There
was one other key thing they knew.
They knew that these genes were in
the nucleus.  And I'll tell you the
full story of how even that insight
was arrived at.
But I'll just show you for the
moment this little movie.
This shows some chromosomes that
are all bunched up and are just
pulled apart at the time of the cell
division.  Those chromosomes,
as we now know, are made of DNA.
But in essence what people had seen
through the microscope was these
chromosomes or colored things that
they could stain.
They could sort of see something
had doubled.  And just before the
cell divided the two sets separated
and each cell got a new set.
So that's about what people knew.
They had those properties.  They're
in the nucleus.
They knew about as much as you do.
They knew the major classes of
biomolecules in a cell.
So what do you think you would need
to do to show that DNA is the
genetic material,
encodes the genes?
Find somebody near you.
I'll give you a minute or so.
I'd like to hear what kind of ideas
you come up with.
Then I'll tell you how it happened.
But I want to hear.  Why don't you
think about it and just see if you
can come up with a couple ideas for
me, what you'd need to figure out.
Well, let's just see what kind of
ideas anybody got.
If you wanted to make me believe
that DNA is doing that,
or I think it's a protein for the
moment, that's what I think is most
likely, but what you do think?
Anybody got an idea?
Mess up the DNA and see if we can
mess up the cell.
How are we going to do that?
I can break a cell open and I guess
I can purify DNA and I can analyze
it.  And it's got four bases in it
and it's got sugars and phosphates.
At that point nobody could sequence
DNA.  We didn't even
know the structure.
Yeah?  Take it out of one cell and
put it into another.
And what would you expect to happen
then?
OK.  That's a really nice idea.
Somehow if you took the DNA and
moved it from one cell to another
that the characteristic of this cell
would be somehow carried
over.  OK.
That's in fact the way it happened
but not as simply as that,
as I'll tell you, but that's exactly
the essence of it.
One little problem.
Maybe we'll see if anybody has a
thought on this.
If I purify DNA,
I mean nothing's ever really pure,
right?  You get it out and there's
always little bits of stuff.
And someone can always argue,
well, yeah, it's 99% DNA, but it's
the other bits you cannot
get rid of.  Yeah?
If you used radioactive material,
how is that going to help us?
It does contain nitrogen.
Well, it gets a little complicated.
Certainly nucleic acids have like
phosphate in them, but so does RNA.
That's going to be hard.
Maybe if I had a mixture of things
and I wanted to prove whether
something was let's say DNA,
a protein or something, you need
some really specific way of saying I
did something to the DNA and not to
the protein or something like that.
Have you heard anything in this
course that is really specific?
Enzymes.  Do you think that'd give
you an idea of how you might do it?
If I've got a tube and it's mostly
DNA and maybe a bit of protein and
something, and let's say his idea is
working, that we can take the DNA
from this cell and put it over into
the second cell and see the
characteristic change,
if I wanted to do something to show
that it was the DNA in the tube that
was responsible,
could you use an enzyme?
And what kind of enzyme would you
want?  Well, what characteristics
would you like it to have?
Nature's probably made it for you
already.  Something that synthesizes
DNA?  Something that breaks down DNA?
Say I treated this tube with some
kind of enzyme and then I wanted to
see the outcome,
what would we want,
an enzyme that did what?
Anybody else got an idea?
You're asserting that it's the DNA
in my prep, I like your idea,
but I need to prove it so I need to
do something to show that it's
actually the DNA and not the other
stuff.  So if I had an enzyme that
did what to DNA?
If I broke it down,
yeah.  We could treat it.
And if your idea is right,
we treat the stuff with something
that specifically breaks down DNA it
won't get transferred.
Does that make sense?  OK.
I mean that's a way you could go at
a proof of this.
And, in fact, that's what happened.
But I'm going to quickly tell you
how it actually happened.
And again, you know,
as I say, I'm trying to tell you a
few things that are besides here are
the facts that you need to know on
exam.  There's a bigger picture here
and this is how research goes,
and particularly in an experimental
science such as biology.
The important early work on this
came from a guy who was known as
Frederick Griffith.
He was in London.  He was a
physician.  He was working
in the 1920s.
And he was studying pneumonia.
That's an infection of the lungs --
-- by bacteria.
There's more than one kind of
bacterium that will cause pneumonia,
but one of the really important ones
clinically was streptococcus --
-- pneumonia.  So it was a bacterium.
It was given that name.
We all have bacterium on us.
I think I told you we have about
ten to the twelfth on our skin,
for example.  And if streptococcus
is on your skin it's not a problem,
but if it gets into your lungs it's
a problem.  And so to live with all
these bacteria with us our
bodies have defenses.
So we have this immune system,
we'll talk about more, and a bunch
of defender cells.
Things that you know as white blood
cells are defenders.
Let's just see here.
I'm going to show you this little
movie.  This is one of your white
blood cells, a special kind of white
blood cell.  That little thing it's
chasing is a bacterium.
These round things are red blood
cells.  I mean doesn't it look like
a dog going after a mouse,
or a cat going after something?
It's chasing it.
It can tell it's there.
This is remarkable.  And it's a
little pixilated,
but this is real.  It's going to
catch it right about there.
And it eats it.  I mean we have
these cells inside us.
That's why you don't die even
though we live in a world that's
surrounded by bacteria.
OK, so we'll go on.  So getting
pneumonia in those days was
a really bad thing.
You get infected,
you get this in your lungs,
and then you have four to six days
this.  So that's the bacterium.
of high fever,
and then the patient would reach
Well, it turns out that
streptococcus is a bacteria like
what's termed as a ìcrisisî.
And one of two things would happen.
They'd either live or they'd die.
And that was it.
I mean this was no fun if somebody
you knew had it because you didn't
know the outcome.
And the outcome wasn't necessarily
437
00:34:27,000 --> 00:34:23,000
very good.  Now you call up the
doctor and they pump you full of
438
00:34:23,000 --> 00:34:19,000
antibiotics, but antibiotics hadn't
been discovered yet.
439
00:34:19,000 --> 00:34:15,000
So this was pretty serious business,
and people were trying to understand
440
00:34:15,000 --> 00:34:11,000
what was happening.
But what was going on during these
441
00:34:11,000 --> 00:34:07,000
four to six days that then led to
one of these two outcomes?
And it has around it something known
as a capsule.  And what that capsule
is polysaccharide.
Remember back to the second lecture
when I was confusing you all by
showing you how sugars could hook
together in all a manner of
different ways?
Well, that's what polysaccharides
are.  You just hook a bunch
of sugars together.
And for this course you don't have
to remember the linkages in
particular.  You just have to
understand that there are different
kinds of linkages,
and every time you hook at it in a
different way you get a different
kind of polysaccharide out of it.
But anyway, the bacterium make this
capsule of polysaccharide.
And it's full of hydroxyl groups
from all those sugars so it attracts
a lot of water around it.
And what it does is it causes a
problem for those defender cells
that we just saw.
Those would be, for example,
a macrophagic kind of white blood
cell.
And it cannot eat something that's
got the capsule.
Now, here's a picture of one of
these capsules on one of these kinds
of bacteria.  You can sort of see it
out here.  It's polysaccharide.
That's the main part of the
bacterium.  And here's another
pixilated thing of one of these
white blood cells eating a bacterium
that doesn't have a capsule.
But watch what happens if the
bacterium has a capsule.
It cannot get a hold of it.
It just cannot quite grab hold of
it.  So what happened during those
days, though, was this capsule which
is a foreign entity to your body
gets recognized in your
immune system.
And your immune system made
antibodies that could recognize that.
We'll talk about what these are,
but all you need to know for the
moment is that they're proteins and
they can be tuned to recognize some
chemical entity with a very,
very high degree of specificity.
So what the body was doing during
this thing was trying to make
antibodies that would help it
recognize this capsule.
And then it decorates the capsule
with these things.
And once it puts antibodies stuck
all over the surface now it can get
a hold of it.  And,
again, a fact you don't have to know.
This whole process is called
opsonization.  The reason they use
the word opsin because opsin is the
Greek word for seasoning.
And it was as if these white blood
cells liked to have their bacteria
seasoned correctly before
they can eat them.
And what's really going on is that
they're decorating them with
antibodies.  So what was going on
after a person got sick,
it was a race between their immune
system trying to make antibodies
which would let their immune system
suppress the infection and the
bacterium which is replicating
unchecked for the first few days.
And that's why it was such a scary
business, because you didn't know
what the outcome was and things
could tip it one way
or the other.
Well, this did suggest a kind of
therapy.  The kind of therapy would
be to isolate a capsule
to inject a horse.
Get the antibodies from the horse.
Why a horse?  A horse is huge,
right?  It makes a lot
of antibodies.
A lot better than injecting a mouse
if you want to get antibodies.
So get antibodies and then inject
the patient.  It's a good idea in
principle.  So you're sort of
short-circuiting this whole process.
The problem was there were more
than 20 kinds of capsules.
And so what people had to do was
they had to isolate
the bacterium --
-- from the patient,
determine the type of capsule.
Let's say it's sort of from capsule
1 up to capsule type 20,
which one it was, and then inject
the correct antibody.
So this was nerve-racking because
it took a while for the bacteria to
grow so it was a pretty tight time
window.  And if you saw the patient
right away that's good,
but if they were partway down the
infection not so good.
So the one other thing to do this,
they didn't bother all the way to
isolate the capsule.
What they would usually do is use
heat-killed bacteria and then you'd
have the capsule and everything.
The bacterium is dead, it cannot do
anything, and they'd inject the
horse with that.
And that would get you the
antibodies with the capsule.
So what Griffith was doing was he
was fiddling around
with this system.
And there was one other discovery
that he made.  Perhaps it wouldn't
surprise you that since the bacteria
surrounded by a molecule absorbs
water that the capsules would look
sort of glistening.
They absorbed a lot of water.
You can see how they look here.
So what they discovered was if they
have a capsule you get what are
called smooth colonies.
And the word colony in this thing
just refers to it started out as one
bacterium and it kept dividing and
dividing and dividing.
And maybe there are ten to the
eighth or ten to the ninth bacteria
in that little colony.
But you can see it.  They've all
got capsules on the outside so it
attracts a lot of water and it looks
wet.  And those are what you saw.
But what they found is if you
waited or grew the cultures up that
you would see some things that
looked dry or they called them rough.
And these turned out to be bacteria
that lacked a capsule.
And so if you might start with a
smooth strain S here and then
isolate from it a rough strain it
might designate it in that kind of
way.  So this is the sort of thing
that Griffith was fooling around
with.  So he started with doing this
kind of experiment.
He took a smooth strain making a
capsule type two,
OK?  So he was injecting a mouse
with this.  And what happened was
the mouse was dead.
This was a virulent form of the
bacterium.  So if he took the rough
mutant, injected the mouse,
the mouse is alive and you saw why.
If it doesn't have a capsule,
the mouse has defender cells and
white blood cells could eat it.
Then he had heat-killed S3.  So
this was a strain of streptococcus
that had a different capsule,
a type 3 capsule, but it was
heat-killed.  Why was he working
with heat-killed stuff?
Because that's what you injected
the horses with to get it.
So what do you think would happen
here?
Since the bacteria are dead,
probably not a big surprise the
mouse is alive.
Now, I don't know whether he did
this on purpose or he did it as a
control, but what he did was he
injected at the same time then R2
plus heat-killed S3.
So he's got two things that don't
do anything, he injects a mouse,
and uh-oh, the mouse dies.
That is a weird result.
That is actually also, though,
the first really key step to
understanding that DNA is a genetic
material.  It doesn't look like it
at this point probably,
but it was.  This is how we learned
this really enormous fact from these
experiments.  He wasn't trying
to figure it out.
He was trying to work out something
else, as you can see,
but it was a bizarre finding.
So what would you think?  I've put
in something that used to have a R2
capsule.  So did it get rejuvenated
somehow by this heat-killed thing or,
as you'd suggested,
did some characteristic get
transferred from here or whatever?
So he isolated the bacteria out of
this, and what he found now was he
had a live bacterium
that was making S3.
So something had been transferred
from this set of dead bacteria into
bacteria that were alive,
and the characteristic had been
passed from the dead bacterium to
the new bacterium,
the other bacterium.
So this is about what Griffith did,
but this problem was picked up by a
scientist at Rockefeller,
Oswald Avery who worked as part of a
team.
And he took this finding and started
to work on it and tried to figure
out, because he saw in this result a
way of finding out what was the
genetic material because somehow
what was in that heat-killed S3 was
the stuff that would transfer
genetic information into another
bacterium.  So he made one
really big discovery.
And that was you didn't need the
mouse at all.  All that was
happening was the mouse,
by dying, was in essence selecting
for smooth bacteria.
So he could simplify things by just
taking a rough bacteria,
taking the heat-killed extract,
putting it in, and now he'd just
look for smooth colonies.
Didn't need any mice at all.
So he was able to see the
characteristic of the capsule being
transferred from some kind of
heat-killed mess of things into a
rough bacterium and changing it into
a live bacterium.
So he started fractionating,
and he did exactly the kind of
this was called transformation.
approach that you suggested.
And he purified and he purified and
matters in the thing was taking DNA
and putting it in.
he purified using as his assay this
ability to pass on this smooth
characteristic.
And what he ended up with was
virtually pure DNA but,
as I said, you know, always never
quite pure.  And somebody can always
argue, well, you've got a little bit
of something else in there.
So he did a really key experiment
and he treated with DNAs,
your experiment.
586
00:46:33,000 --> 00:46:00,000
And it lost the transforming
activity.  So this process of doing
plasma and you stick it into E.
coli so you can grow it up,  that
Initially it described that
phenomenon.  Now that we know what
So if you do a UROP somewhere here,
you clone a piece of DNA into a
process of taking the naked DNA and
putting it inside the bacteria,
you'll call it transformation.
Now, in fact, this result wasn't
accepted right away.
This was published in 1944.
And the general realization that
DNA was the genetic material really
didn't come until the ë50s.
Yet this result proved it, if you
will, but part of the problem was
the world wasn't yet ready to accept
that DNA was a genetic material.
And maybe you can see the problem.
It looked like a monotonous
molecule.  It only had four things
that were different in it.
And if you isolated they were all
often kind of there and about the
same amount.  People thought it was
just an analyst GATC.
It didn't sound like anything had
encoded information.
Proteins looked really attractive.
Twenty different amino acids that
all had different characteristics,
so that was a great place for
storing information.
So the world wasn't quite ready to
accept it, even though the
experimental evidence was there.
And so the result came later.
Now, the last thing I just want to
show you, because there's a kind of
direct link from that Avery
experiment to you guys because a
year or two ago it was the 50th
anniversary of the discovery of DNA.
And Avery worked with a team of two
other people called MacLeod and
McCarty.  This was at the 50th
anniversary, the meeting down at
Cold Spring Harbor celebrating it.
McCarty was the only member of the
team alive.  There he was.
I asked him to autograph my program.
There was his signature.
He just died a little while ago,
and so there's no living connection
to that anymore,
but I have a picture to show you
guys that takes you back from that
experiment to his signature right
there.  OK?  So I'll tell you some
more stuff next lecture.
Have a good weekend.
