Lecture 05:The Founders of Genetic Engineering

1. Professor 1. Professor Paul BergPaul Berg

DNA + SV40 → Recombination DNADNA + SV40 → Recombination DNA

Brief CV Paul Berg witnessed first-hand the history of

recombinant DNA research and regulation, having been in the forefront of both movements since he was a young man.

He became a Professor of Biochemistry at Stanford University School of Medicine in 1959, when he was 33.

He was elected to be the National Academy of Sciences before he was 40, and he gained early recognition and influence when he delineated the key steps in which DNA produces proteins.

Berg was awarded the Nobel Prize for Chemistry in 1980 for his work with DNA.

Paul Berg Curriculum Vitae Born: Brooklyn, NY, June 30, 1926 ； Education  

1948 B.S., Pennsylvania State University (22y) 1952 Ph.D., Western Reserve University (26y)

Professional Background 1950-52 Predoctoral Research Fellow, National Institutes of Health.

(24) 1952-54 Postdoctoral Research Fellow, American Cancer Society

(26y)Institute of Cyto-physiology, Copenhagen, Denmark Dr. Arthur Kornberg, Washington University School of Medicine, St. Louis, Missouri State

1954 Scholar in Cancer Research, American Cancer Society, Department of Microbiology, Washington University School of Medicine (28y) 1955-59 Assistant to Associate Professor of Microbiology, Washington University School of Medicine (29-33y)

1959-60 Associate Professor of Biochemistry, Stanford

University School of Medicine (33-34y) 1960-69 Professor, Department of Biochemistry, Stanford

University School of Medicine (34-43y) 1969-74 Chairman, Department of Biochemistry, Stanford

University School of Medicine (43-48y) 1970-94 Sam, Lulu and Jack Willson Professor ( 斯坦福大学 终身教授 ) of Biochemistry (44-64y) 1985-2000 Director, Beckman Center for Molecular and

Genetic Medicine (59-74y) 1994-2000 Vivian K. and Robert W. Cahill Professor in

Biochemistry and Cancer Research (68-74y) 2000-pre. Cahill Professor in Biochemistry , Emeritus, and

Director of the Beckman Center for Molecular and

Genetic Medicine, Emeritus (74-82y)

Selected Bibliography 1. Physical and Genetic Characterization of Deletion Mutants of Simian Virus 40

Constructed In Vitro. Charles Cole, Terry Landers, Stephen Goff, Simone Manteuil-Brutlag, and Paul BergPaul Berg. J. Virol., 24: 277 - 294 (1977).

2. A Biochemical Method for Inserting New Genetic Information into SV40 DNA: Circular SV40 DNA Molecules Containing Lambda Phage Genes and the Galactose Operon of E. coli. David A. Jackson, Robert H. Symons, and Paul Paul BergBerg. Proc. Nat. Sci. USA, 69, 2904 (1972).

3. Construction of Hybrid Viruses Containing SV40 and Lambda Phage DNA Segments and Their Propagation in Cultured Monkey Cells. Stephen P. Goff, and Paul BergPaul Berg. Cell, 9:695 (1976).

4. Synthesis of Rabbit -Globin in Cultured Monkey Kidney Cells Following Infection with a SV40 -Globin Recombinant Genome. R.C. Mulligan, B.H. Howard, and Paul BergPaul Berg. Nature, 277, 108 - 114 ( 1979).

5. Expression of a Bacterial Gene in Mammalian Cells. R.C. Mulligan and Paul Paul BergBerg, Science 209, 1422 - 1427 (1980).

CNSCNS

Nobel lecture, 8 December, 1980

Fig. 2. A physical and genetic map of SV40 DNA.

Fig. 4. A scheme for construction ofSV40 transducing genomes in vitro.

Fig. 3. The construction of SV40-λphage recombinant DNA.

Address: 838 Santa Fe Ave., Stanford, CA 94305

2. Prof. Frederick Sanger2. Prof. Frederick Sanger

Protein sequencingProtein sequencingDNA sequencingDNA sequencing

F. Sanger received two Nobel prizes

Frederick SangerThe Nobel Prize in Chemistry 1958 and 1980 Autobiography I was born on 13th August 1918 in the village of

Rendcombe in Gloucestershire, where my father,

also Frederick Sanger, was a medical practitioner.

Influenced by him, I soon became interested in

biology and developed a respect for the importance

of science and the scientific method.

At Bryanston School and St John's College,

Cambridge, I was probably above average but not

an outstanding scholar. Initially I had intended to st

udy medicine, but before going to University I had

decided that I would be better suited to a career in

which I could concentrate my activities and interest

s more on a single goal than appeared to be possib

le in my father's profession.

So I decided to study science and, on arrival at

Cambridge .

I was married to Margaret Joan Howe in 1940. Although not a scientist herself she has contributed more to my work than anyone else by providing a peaceful and happy home. We have two sons, Robin and Peter, born in 1943 and 1946, and a daughter, Sally Joan, born in 1960. Apart from my work my main interests are gardening and what can best be described as “messing about in boats”.

Beautiful Family Life Autobiography

Frederick Sanger received his first Nobel Prize in Chemistry in 1958

Since 1943 his work has been concerned largely with problems related to the determination of the structure of proteins. These studies resulted in the determination of the structure of insulin.

From 1944 to 1951 he held a Beit Memorial Fellowship for Medical Research and since 1951 he has been a member of the External Staff of the Medical Research Council. His present position is Head of the Division of Protein Chemistry in M.R.C. Laboratory for Molecular Biology at Cambridge.

Nobel Lecture, December 11,

1958

In 1962 I moved to their newly built Laboratory of Molecular Biology in Cambridge, together with F.H.C. Crick, J.C. Kendrew, H.E. Huxley and A. Klug. In this atmosphere I soon became interested in nucleic acids. My work on nucleic acids is summarized in my Nobel lecture. This work has not been done single-handed and it owes much to the excellent collaborators I have had.

Most of these have been students and postdoctoral fellows spending a few years in the laboratory and bringing their experience and ideas with them, but I feel particularly indebted to my more permanent colleagues, B.G. Barrell, A.R. Coulson and G.G. Brownlee, who have contributed so much to the methods we have developed.

Didesoxy DNA sequencing The chemical method of DNA

sequencing hat largely been superseded by the method of F. Sanger, which uses ddNTP to terminate enzymiclly synthesized copies of template (Fig. I). A sequencing primer is annealed to a dsDNA template molecule and a DNA polymerase extends the primer using ddNTPs.

The extension reaction is split into four specific ddNTPs and separately with one of the four specific ddNTPs, and S four samples (usually radioactive) are analyzed by PAGE.

DETERMINATION OF NUCLEOTIDESEQUENCES IN DNA

Nobel lecture, 8 December 1980

The term "shotgun" to denote sequencing of a large number of random DNA clones was first coined by F. Sanger and co-workers in 1982.

Thank you !Thank you !