The First
Sequenced Gene
CONRAD GREAVES
The Beginning

Friedrich Miescher

Discovered DNA in 1869

Studied White Blood Cells

Collected Used Bandages
A near by Hospital

After removing Lipids and
Proteins found leftover stuff
Called it Nuclein
Protein Centric Paradigm

For a long time scientists thought that proteins and protein
sequences contained the code of life

DNA was largely ignored until 1944 when Oswald Avery, Colin
MacLeod, and Maclyn McCarty showed that altering the DNA of an
organism can change the traits it expresses.
"then nucleic acids must be regarded as
possessing biological specificity the chemical
basis of which is as yet undetermined.” Avery,
McCarty and Macleod -1944
Frederick Sanger

Driving force behind genetic
sequencing

Determined amino acid
sequence of insulin (1955)

Developed Plus and Minus
sequencing method (1975)

Sequenced first genome - Phi X
174 (1977) Bacteriophage

Developed Sanger sequencing
(1977)
Plus and Minus
Method
Sanger and Clouson (1975)

8 different polymerase reactions

4 missing one nucleotide (AGT, GTC, CGA,
AGC)

4 with only 1 nucleotide (A,G,T,C)

~50 bases

Used to sequence Phi X 174 genome

Not good for homopolymer runs
Phi X 174
Maxam-Gilbert
sequencing (1977)

Similar to Plus and Minus

Useful for homopolymer runs

Uses double stranded DNA

Four Chemical Cleavage
Reactions

A+G ,G, C, C+T

A+G prefers A

C+T prefers T
Chain-termination methods
(Sanger Sequencing)

Dideoxy method

4 reactions setup

Each with a different ddNTP

1% termination rate

Product is electrophoresed in
12% polyacrylamide

Reads od ~100bp

Disovered by Sanger in 1977
The First Sequenced Gene
LAC1

Enzyme in E-Coli

Protein sequence
already known

Sequenced using
Maxam-Gilbert
Method

Published 1978 by
Philip J Farabough

Compared
sequenced DNA
with protein
sequence
References

https://outlook.office.com/owa/?realm=unm.edu#path=/mail/search/attachmentlightbox

https://en.wikipedia.org/wiki/DNA_sequencing

http://www.454.com/downloads/news-events/history-of-genome-sequencing_FINAL.pdf

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2094077/

http://www.mun.ca/biology/scarr/4241_RMC_Sequencing.html

http://www.genetics.org/content/162/2/527.full

http://www.mun.ca/biology/scarr/4241_RMC_Sequencing.html

https://en.wikipedia.org/wiki/Frederick_Sanger

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2094077/

https://en.wikipedia.org/wiki/DNA_sequencing

https://www.youtube.com/watch?v=8kK2zwjRV0M

http://blog.wellcomelibrary.org/2013/05/fred-sanger-a-maker-of-modern-genetics/

https://en.wikipedia.org/wiki/Friedrich_Miescher

http://www.genetics.org/content/162/2/527.full