Contemporary Concepts in Cellular and Molecular Biology for the Medical Physicist

Barry S. Rosenstein, Ph.D.

Mount Sinai School of Medicine NYU School of Medicine
www.che m.ucsb.e du/.../ima ges/Wats onCrick.j pg

Alberts et al. Molecular Biology of the Cell, 1994

Alberts et al. Molecular Biology of the Cell, 1994

Alberts et al. Molecular Biology of the Cell, 1994

Alberts et al. Molecular Biology of the Cell, 1994

Alberts et al. Molecular Biology of the Cell, 1994

Alberts et al. Molecular Biology of the Cell, 1994

Alberts et al. Molecular Biology of the Cell 1994

Lewin, Genes V, 1994

Hall, Radiobiology for the Radiologist, 2000

Lewin, Genes V, 1994

Lewin, Genes V, 1994

VECTORS
RESTRICTION ENDONUCLEASES

Self-replicating DNA molecule that has the ability to carry another foreign DNA molecule into a host cell. Purpose - insert a fragment of DNA into a vector so that it can be replicated and grown into quantities suitable for study.

Alberts et al. Molecular Biology of the Cell, 1994

PLASMID

Alberts et al. Molecular Biology of the Cell, 1994

Alberts et al. Molecular Biology of the Cell, 1994

COMMONLY USED REPORTER GENES

REPORTER GENE ASSAYS

Chloramphenicol Acetyltransferase (CAT) Firefly Luciferase -Galactosidase Secreted Alkaline Phosphatase (SEAP) Human Growth Hormone (hGH) -Glucuronidase (GUS) Green Fluorescent Protein (GFP)

Hall and Giaccia, Radiobiology for the Radiologist, 2006

GENOMIC LIBRARY

LIBRARIES

Compilation of DNA fragments that make up the entire genome.

athena.bioc.u vic.ca/.../copy 13_of_sampl e/phage

cDNA LIBRARY
DNA complementary to mRNA - only the expressed genes of a particular cell.
8e.devbio.com/im ages/ch04/0405fi g2.jpg

EXPRESSION LIBRARY

http://fig.cox.miami.ed u/~cmallery/150/gene/s f16x5.jpg

Alberts et al. Molecular Biology of the Cell, 1994

GEL ELECTROPHORESIS

Hall and Giaccia, Radiobiolog y for the Radiologist, 2006

www.cbs.dtu.dk/staff/dave/roanoke/fig5_33.jpg

http://www.mun.ca/biology/scarr/Lab_5_gel_1999.gif

Hall and Giaccia, Radiobiology for the Radiologist, 2006

SDS POLYACRYLAMIDE GEL ELECTROPHORESIS

http://fig.cox.miami.edu/~cmallery/2 55/255tech/mcb3.32.SDS.elect.jpg Hall and Giaccia, Radiobiology for the Radiologist, 2006

POLYMERASE CHAIN REACTION

Alberts et al. Molecular Biology of the Cell, 1994

Alberts et al. Molecular Biology of the Cell, 1994

REVERSE TRANSCRIPTASE PCR

RT PCR

qPCR Using Real-Time PCR

https://www. finnzymes.fi/ realtime_qpc r/dynamo_ca pillary_sybr_ green_qpcr_ kit.html

NORTHEN BLOT

Southern Blotting DNA Northern Blotting RNA

Alberts et al. Molecular Biology of the Cell, 1994

http://www.mcmaster.ca/inabis98/higuchi/noda0326/f3small.gif

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FAR-WESTERN BLOTTING

WESTERN BLOT

http://www.bseinqu iry.gov.uk/report/vo lume2/fig1_8.htm

http://www.bioon.com/experiment/UploadFiles/200408/20040803131929294.gif

EXPRESSION MICROARRAYS

www.Affymetrix.com

www.Affymetrix.com

11

www.Affymetri x.com

www.Affymetr ix.com

www.Affyme trix.com

www.Affymetrix.com

12

www.Aff ymetrix.c om

Hall and Giaccia, Radiobiology for the Radiologist, 2006

SINGLE NUCLEOTIDE POLYMORPHISM

SNP
http://ww w.ogt.co. uk/image s/humangeneexpressio n.gif www.seq.ubc.ca/deco de.ne-icelands-dna/

Bruchova. Leukemia Res.28:1,2004

13

PERSONALIZED MEDICINE
The use of detailed information about a patient's genotype in order to select a medication, therapy or preventative measure that is particularly suited specifically to that patient.

www.sanger.ac.uk

SINGLE-STRANDED CONFORMATION POLYMORPHISM SSCP

SNP ASSAYS

National.diagnostics.combined/images/2_3_2/gif

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MOLECULAR BEACONS
5 Nuclease TaqMan Assay

Clinical Science www.clinsci.org. Clin. www.clinsci.org. Clin. Sci. (2005) 109, 365Sci. 365379

www.ebiotra de.com/,,,/fig. molecular_be acons.gif

SNP/CNP (Genotyping) Arrays

Affymetrix

Illumina
www.Affymetrix.com

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http://www. accessexc ellence.org /RC/VL/GG/ transgenic. php

http://www.biken.osaka-u.ac.jp/act/act_okabe_e.php

Enzyme-Linked ImmunoSorbent Assay

KNOCKOUT MICE

http://www.niaaa.nih.g ov/Resources/Graphics Gallery/Genetics/homa n.htm

http://64.202.120.86/upload/image/articles/2006/biopen/biopen-elisa-schematic.jpg

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Electrophoretic Mobility Shift Assay EMSA

IMMUNOPRECIPITATION

www.biochem.arizona.edu/.../Am64./8a.gif

www.molecularstation.com

CHROMATIN IMMUNOPRECIPITATION

ChIP

ChIP-on-chip

http://en.wikipedia.org /wiki/ChIP-on-chip www.vinicibiochem.it/images/chip_flow.jpg

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DNase I Footprinting

Alberts et al. Molecular Biology of the Cell, 1994

Alberts et al. Molecular Biology of the Cell, 1994

siRNA

miRNA

www.scq.ubc.ca/wpcontent/siRNA.gif

www.microrna.ic .cz/obr/image003 .png

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PULSED FIELD GEL ELECTROPHORESIS

DNA STRAND BREAK ASSAYS

Herschleb et al., Nature Protocols 2: 677, 2007

COMET ASSAY

www.scielo.br/img/fbpe/gmb/v23n1/2339f1.jpg Burdak-Rothkamm Oncogene 26: 993, 2007

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http://jamescbende r.com/bendersblog /Default.aspx

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