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7/11/2011

The Central Dogma of Molecular Biology: From DNA to RNA to Proteins

MBB 1 July 5, 2011

http://www.pub.ac.za

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Cell Nucleus
Chromosome

DNA

DNA Packaging:

www.csu.edu.au/.../molbol/images/8_10.jpg

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DNA, RNA and Proteins

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http://www.ncbe.reading.ac.uk/DNA50/Resources/wc1993.gif

DNA Structure

We have discovered the secret of life. Francis Crick

DNA

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Nitrogenous Bases

http://www.ca.uky.edu/Agripedia/glossary/agriart/

Nitrogenous Bases

Purines

Pyrimidines

http://www.ca.uky.edu/Agripedia/glossary/agriart/

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Nucleotides

http://ghs.gresham.k12.or.us/science/ps/sci/ibbio/chem/notes/chpt14/nucleotide1.gif

http://bvc.kribb.re.kr/download/image/nucleotide2.gif

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http://www.emc.maricopa.edu/faculty/farabee/biobk/BP2.gif

RNA

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Types of RNA
messenger RNA (mRNA): Carries the genetic information out of the nucleus into cytoplasm for protein synthesis ribosomal RNA (rRNA): Makes up ribosome, constitutes 50% of a ribosome, which is a molecular assembly involved in protein synthesis transfer RNA (tRNA): Decodes the information in mRNA

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http://scienceblogs.com/clock/upload/2006/11/a2%20DNA-RNA-ribosome2.jpg

Proteins

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Protein Buliding Blocks: Amino Acids

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Cellular Proteins
Enzymes Catalyze chemical reactions Cytoskeletal proteins Give cell its shape and motility Receptor proteins Bind to molecular stimuli from outside the cell Signal transduction Relaying of signal into intracellular events Perform almost every other function in a cell

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Sickle Cell Anemia

Normal red blood cells Sickled RBC

Sickle Cell Anemia

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The Central Dogma of Molecular Biology: From Genes to Proteins

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"My mind was, that a dogma was an idea for which there was no reasonable evidence. You see?!" And Crick gave a roar of delight. "I just didn't know what dogma meant. And I could just as well have called it the 'Central Hypothesis,' or you know. Which is what I meant to say. Dogma was just a catch phrase. - Horace Freeland Judson, The Eighth Day of Creation

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From Genes to Proteins

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RNA transcription: From DNA to RNA

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Messenger RNA
mRNA Copies information from DNA to bring out into cytoplasm Is used by transfer RNA (tRNA) and ribosomes to produce protein Is made up of codons as informational units

Codon

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Genetic Code
the letters of the genetic alphabet... are the nucleotides A, T, G, & C of DNA unit of information is CODON = genetic 'word' 3 nucleotides = 1 codon (word) = 1 amino acid Amino acids are the building blocks of proteins Amino acid : word :: protein : sentence

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Stop codons mark the end of the protein Amino acid : word Protein : sentence

www.accessexcellence.org

Stop codon : period

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Translation: From RNA to Protein

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Working with DNA


MBB 1 July 7, 2011

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Advances in DNA Technology


1995 entire genome of Haemophilus influenzae was sequenced 2003 human genome was completed (3 billion base pairs) 2007 hundreds of prokaryotic genomes and a dozen of eukaryotic genomes

Recombinant DNA Technology

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Recombinant DNA
DNA molecules formed when segments of DNA from two different sources often different species are combined in vitro (in a test tube) Central to genetic engineering
Direct manipulation of genes for practical purposes

DNA/gene cloning
The production of multiple copies of a single gene

Recombinant DNA: Gene Cloning


The entire DNA molecule is very long Contains thousands of genes Scientists need to work directly with specific genes Multiple identical copies of a single gene are often needed

GENE CLONING!!!

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Manipulating DNA: Ingredients


Restriction enzymes (a.k.a. restriction endonucleases)
Cut DNA on specific sites (restriction sites)

Ligase
Joins DNA strands together

Plasmid DNA
Circular bacterial DNA (replicate separately from bacterial DNA)

Gene/DNA of interest
Foreign gene that is introduced into the plasmid DNA and evetually the bacterium

Bacteria (E. coli)


where you will insert the plasmid DNA; bacteria is allowed to grow to make many copies of the gene of interest

Use of Restriction Enzymes

http://fig.cox.miami.edu/~cmallery/150/gene/sf15x1box.jpg

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Plasmid DNA
Double stranded, circular Separate from bacterial DNA Can replicate independently Carries accessory genes antibiotic resistance, toxins, etc.

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Recombinant DNA Technology


Transformation change in genotype and phenotype due to the integration of external DNA by the cell

Uses of Gene Cloning

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Visualizing DNA

Visualizing DNA

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Visualizing DNA

Sequencing DNA: DNA extension

http://seqcore.brcf.med.umich.edu/doc/educ/dnapr/sequencing.html

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Sequencing DNA: Using dideoxynucleotides

http://seqcore.brcf.med.umich.edu/doc/educ/dnapr/sequencing.html

Sanger Method

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Automated Sequencing

Sequencing DNA: Reading the sequence using a computer

http://seqcore.brcf.med.umich.edu/doc/educ/dnapr/sequencing.html

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Polymerase Chain Reaction


Kary B. Mullis, working for Cetus Corporation, developed the Polymerase Chain Reaction in 1983. The technique allows for the rapid synthesis of DNA fragments. In about an hour, over 1 million copies of a DNA strand can be made Mullis left Cetus in 1986. For his development of PCR, he was co-awarded the Nobel Prize in chemistry in 1993. Mullis is currently doing HIV and AIDS research The technique has been invaluable to the development of biotechnology and genetic engineering. Its real world applications include gene mapping and forensic science.

Kary Mullis: Surfer-Scientist

www.revolt.it

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PCR

library.thinkquest.org

PCR

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PCR

library.thinkquest.org

PCR

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