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Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

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Page 1: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.
Page 2: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Welcome toMolecular Biology Through Discovery

Tuesday, 17 September 2013DNA Structure / Benzer (1959)Finally found a mentor!

Is there anything that we should physically or

electronically be turning in as proof?

Page 3: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

DNA Structure

ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ...

A class discussion on the tetranucleotide

hypothesis might help.

Page 4: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

DNA Structure

ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ...TAGGCCACTGCCCAACCCTCCATCATAAAACTTGGGCTTGGGAGGCAGAGCCTAACCTCTCTCACTCTAGACAGGTCTAAGATGATTGGGAACGAAATGAGCCGTCTCGACTTTTTCGCGAAGTGGCTAA ...

#1: Hello!

#2: Complete description of our civilization

Page 5: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

DNA Structure

Page 6: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

DNA StructureSQ7. What would you expect to happen to the diffraction pattern if the helix shown in Fig. 8B-D were squashed, i.e. the helix were more tightly wound, more like a solenoid?

Page 7: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

DNA Directionality & Palindromes

…would like to go into deeper discussion about palindromic sequences

Page 8: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Palindromic Sequences

What is it?

What about with DNA? AGTTGA

Backwards = forwards ROTATOR

TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

• DNA is double stranded

SQ16. … why is AGTTGA not a DNA palindrome?

Page 9: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

What is it?

What about with DNA? AGTTGA

Backwards = forwards ROTATOR

• DNA is redundant

TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

• DNA is double stranded

Palindromic Sequences

Page 10: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

What is it?

What about with DNA? AGTTGA

Backwards = forwards ROTATOR

• DNA is redundant

TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

• DNA is double stranded

TTAATGTGAGTTAGCTCACTCATTAATGAGTGAGCTAACTCACATTAA

• DNA has direction (read 5’->3’)

5’- -3’3’- -5’

Palindromic Sequences

Page 11: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

5’- -3’3’- -5’

TAT GGCATGCTAGC

TTAAT TCATTAATTA AGTAA

CGTACGATCGG TAT

DNA: cruciform

RNA: stem/loop

Palindromic SequencesPalindromic sequences as structural RNAThe full biological roles of palindromic

sequences is still a bit unclear to me

SQ18. What is the relationship between a palindromic sequence and a potential hairpin structure?

Page 12: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

5’- -3’3’- -5’

tRNA

UAU GGCAUGCUAGC

UUAAU UCAUU

DNA: cruciform

RNA: stem/loop

Palindromic SequencesPalindromic sequences as structural RNA

Page 13: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

recognizes GTGAGTT

NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

NNNNNNNNNNNNNNNNNNNNNNNNNN

TTAATGTGAGTTAGCTCACTCATT AATGAGTGAGCTAACTCACATTAA

Palindromic SequencesPalindromic sequences as protein binding sites

Page 14: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

NNNNNNNNNNNNNNNNNNNNNNNNNN

TTAATGTGAGTTAGCTCACTCATT AATGAGTGAGCTAACTCACATTAA

Palindromic SequencesPalindromic sequences as protein binding sites

Page 15: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Palindromic Sequences

Palindromic sequences as protein binding sites

NNNNNNNNNNNNNNN

NNNNNNNNNNNNNNN

NNNNNNNNNNNNNNNNNNNNNNNNNN

TTAATGTGAGTTAGCTCACTCATT AATGAGTG

AGCTAACT

CACATTAA

Page 16: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Palindromic Sequences

Palindromic sequences as protein binding sites

NNNNNNNNNNNNNNN

NNNNNNNNNNNNNNN

NNNNNNNNNNNNN

NNNNNNNNNNNNN

TTAATGTGAGTTAGCTCACTCATT

AATGAGTGAGCTAACTCACATTAA

Page 17: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Palindrom

ic Sequences

Palindromic sequences as protein binding sites

NNNNNNNNNNNNNNN

NNNNNNNNNNNNNNN

NNNNNNNNNNNNN

NNNNNNNNNNNNN

TTAATGTGAGTTAGCTCACTCATT

AATGAGTGAGCTAACTCACATTAA

Page 18: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

recognizes GTGAGTT

Palindromic Sequences

Palindromic sequences as protein binding sites

NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

NNNNNNNNNNNNNNNNNNNNNNNNNN

TTAATGTGAGTTAGCTCACTCATTAATGAGTGAGCTAACTCACATTAA

Page 19: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Palindromic Sequences

Palindromic sequences as protein binding sites

NNNNNNNN

NNNNNNN

NNNNNNNN

NNNNNNN

NNNNNNNN

NNNNN

NNNNNNNN

NNNNN

TTAATGTG

AGTTAGCT

CACTCATT

AATGAGTGAGCTAACTCACATTAA

Page 20: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Palin

drom

ic Se

quen

ces

Palin

drom

ic se

quen

ces a

s pro

tein

bind

ing

sites

NNNNNNNNNNNNNNN

NNNNNNNNNNNNNNN

NNNNNNNNNNNNN

NNNNNNNNNNNNN

TTAATGTGAGTTAGCTCACTCATT

AATGAGTGAGCTAACTCACATTAA

Page 21: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Palindromes: Serve as binding sites for dimeric protein

Palindromic SequencesPalindromic sequences as protein binding sites

NNNNNNNNNNNNNNNNNNNNNNNNNNNNNN

NNNNNNNNNNNNNNNNNNNNNNNNNN

TTAATGTGAGTTAGCTCACTCATT AATGAGTGAGCTAACTCACATTAA

Page 22: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

GTA ..(8).. TAC

5’-GTA ..(8).. TACNNNNNNNNNNTANNNTNNNNNNNNNNNNNNNNNNNNNNNNNNNNATGNNNNNNNNNNNNNNNN3’-CAT ..(8).. ATGNNNNNNNNNNATNNNANNNNNNNNNNNNNNNNNNNNNNNNNNNNTACNNNNNNNNNNNNNNNN

gene

Palindromic SequencesPalindromic sequences as protein binding sites

Page 23: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

GTA ..(8).. TAC

5’-GTA ..(8).. TACNNNNNNNNNNTANNNTNNNNNNNNNNNNNNNNNNNNNNNNNNNNATGNNNNNNNNNNNNNNNN3’-CAT ..(8).. ATGNNNNNNNNNNATNNNANNNNNNNNNNNNNNNNNNNNNNNNNNNNTACNNNNNNNNNNNNNNNN

gene

Transcription factor

RNA Polymerase

Palindromic SequencesPalindromic sequences as protein binding sites

RNA

TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

5’- -3’3’- -5’

How many 12-nt palindromes would you expect at random in the genome of Anabaena variabilis?

Page 24: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.
Page 25: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Probability

How often A-A-A-A-A-A?

Page 26: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Probability

How often 1-1-1-1-1-1?

Page 27: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Probability

How often A-A-A-A-A-A?

Page 28: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Probability

How often 1-1-1-1-1-1?

Page 29: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Probability

How often 1-2-1-1-1-1?

Page 30: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Probability

How often 1-1-1-1-1-1?

Page 31: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Probability

How often 1-1-1-1-1-1?

Page 32: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Probability

How often A-A-A-A-A-A?

Page 33: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Probability

How often A-T-G-G-T-A?

Page 34: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Probability

How often A-T-G-G-T-A?

Counting

Page 35: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

GTA ..(8).. TAC

5’-GTA ..(8).. TACNNNNNNNNNNTANNNTNNNNNNNNNNNNNNNNNNNNNNNNNNNNATGNNNNNNNNNNNNNNNN3’-CAT ..(8).. ATGNNNNNNNNNNATNNNANNNNNNNNNNNNNNNNNNNNNNNNNNNNTACNNNNNNNNNNNNNNNN

gene

Transcription factor

Palindromic SequencesPalindromic sequences as protein binding sites

RNA

TTAATGTGAGTTAGCTCACTCATTAATTACACTCAATCGAGTGAGTAA

5’- -3’3’- -5’

How many 12-nt palindromes would you expect at random in the genome of Anabaena variabilis?

Page 36: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Welcome toMolecular Biology Through Discovery

Tuesday, 17 September 2013DNA Structure / Benzer (1959)

Page 37: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Fermi Problems

What is the question?

Page 38: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Fermi Problems

• Focus on what is being asked

• Estimate what can be estimated

• Set limits on what can't

How many piano tuners are there in Chicago?

Page 39: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Fermi Problems

• Focus on what is being asked

• Estimate what can be estimated

• Set limits on what can't

What fraction of the cells in your body are bacterial?

Page 40: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Fermi Problems

What is the question?What do you need?

Page 41: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

Hoping you will go over the experiment. It is so hard to

visualize what goes down in a lab without actually laying on hands.

Page 42: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Wrinch (1936) – What is a Gene?

Morgan T (1934) Nobel Lecture

Drosophilachromosome

What are genes?... There is no consensus of opinion amongst

geneticists as to what the genes are – whether they are

real or purely fictitious...

Page 43: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

Page 44: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

Page 45: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

Complementation Test

Page 46: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

Complementation Test

Phage

E. coli

Page 47: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

Page 48: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

Recomination Test

Phage

E. coli

(Cis-Trans Test)

Measuredistance

Page 49: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

Recomination Test

Phage

E. coli

(Cis-Trans Test)

Measuredistance

Page 50: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

Complementation Test?

Phage

E. coli

Recombination Test?

Page 51: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

Whatis a

Gene?

SQ2. Is there any reason to believe from classical genetics that genes have internal structure?

Page 52: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

rII region of bacteriophage T4

What do you need for the experiment?

Page 53: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

rII region of bacteriophage T4

mutation

Page 54: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959) experiment

Page 55: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959) experiment

Page 56: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959) experiment

Page 57: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959) experiment

Page 58: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959) experiment

Page 59: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959) experiment

Page 60: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Problem Set 5

Page 61: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Problem Set 5

Page 62: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)

rII region of bacteriophage T4

Page 63: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)Determine distances between markers by

Recombination

Page 64: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)I still have trouble connecting recombination with the distance b/w genetic markers.

Page 65: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)I still have trouble connecting recombination with the distance b/w genetic markers.

Richard Twyman, University of Warwick

Page 66: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Benzer (1959)I still have trouble connecting recombination with the distance b/w genetic markers.

Elizabeth Ellis University of Strathclyde

Page 67: Welcome to Molecular Biology Through Discovery Tuesday, 17 September 2013 DNA Structure / Benzer (1959) Finally found a mentor! Is there anything that.

Goodbye fromMolecular Biology Through Discovery

Thursday, 19 September 2013

ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ACTG ...G-O-O-D-B-Y-E