Introduction to Basic Molecular Biology

Dr. Ram Krishna Yadav

Agriculture and Forestry University (AFU)

Rampur, Chitwan

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http://www.promocell.com

The explosion of knowledge brought about by improvements in microscopy, biochemistry, and genetics has led to a depth of understanding of cell structure and function undreamed of by the earliest cell biologists.

Learning Objectives:

• Knowledge and understanding in cell and molecular biology research in Livestock, Fishery and others
• Gain independent thought into research regarding Molecular level.
• Encourage the new young scientists of tomorrow.

Biology is fabulous!

Can you think of any cell and molecular biology words that come to mind?

• Write down key words

Lets share them together.

Actin Filaments Animal cells Plant cells gene

Plasma Membrane

Extracellular Matrix

Golgi Apparatus somatic cell

Lysosome mitosis DNA replication mitosis Centrioles

Ribosomes Prokaryotes Eukaryotes Apoptosis chromatin fiber

Centrosomes

Mitochondria transgene Microinjection

Rough Endoplasmic Reticulum Genoptype

Smooth Endoplasmic Reticulum

Nucleus

DNA methylation

Histones epigenetics anaphase

Cell Cycle (G0, G1, M and G2 phases) Genetic finger printing

Prophase, Metaphase, Anaphase and Telophase

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There are some many………

Genetics, Organelles, Cells, Tissues and Organs

Complexity

all are amazing because they are all made up of about 37 trillion cells of 200 different types.

• Cells are the fundamental unit of living organisms (the building blocks)
• The invention of the telescope made the Cosmos accessible to human observation - the microscope opened up smaller worlds.
• Cell biology has been studied since 16^th century
• Physiological properties
• Structure
• Organelles
• Life cycle
• Division – mitosis and meiosis
• Cell death
• Interactions between cells an their extracellular environment
• Core for developmental biology, stem cell research, immunology.

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Cell Biology

• All known living things are made up of cells.
• The cell is structural & functional unit of all living things.
• All cells come from pre-existing cells by division (spontaneous generation does not occur).
• Cells contain hereditary information which is passed from cell to cell during cell division.
• Cells are similar in chemical composition.
• All energy flow (metabolism & biochemistry) of life occurs within cells.

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References: Mullock BM & Luzio JP, Theory of Organelle Biogenesis: a Historical Perspective (2005). Landes Biosciences, P. 1-13.

Muzzarello P, A Unifying Concept: the History of Cell Theory (1999). Nature Cell Biology, 1, E13-E15.

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Modern Cell Theory

What is molecular biology?

• The attempt to understand biological phenomena in molecular terms
• The study of gene structure and function at the molecular level
• As a result, It is the study of molecular basic of the process of replication, transcription and translation of the genetic material.
• Molecular biology mainly concerns itself with
• Understanding of interactions between the various systems of a cell, including the interactions between DNA, RNA and protein biosynthesis
• learning how these interactions are regulated.

Molecular Biology

• This field overlaps with other areas of biology and chemistry, particularly genetics and biochemistry.
• It is the joining of aspects between genetics and biochemistry

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Components Involve in Molecular Biology�All Life depends on 3 critical molecules

DNA

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RNA

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Protein

Structure

Static

Function

Dynamic

Function

Dynamic

Inside a Living Cell�

A Brief History

• Since the late 1950s and early 1960s, molecular biologists have learned to
• Characterize, isolate, and manipulate the molecular components of cells and organisms, which are:
• DNA, the storage of genetic information
• RNA
• Proteins, the major structural and enzymatic type of molecule in cells.

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1595 – Jansen: developed the first light microscope�1655 – Hooke: described ‘cells’ in cork.�1833 – Brown: described the cell’s nucleus from the orchid.�1839 – Schleiden & Schwann: proposed cell theory (all organisms are comprised of cells).�1858 – Rudolf Virchow: omnis cellula e cellula - cells develop only from pre-existing cells by a process called cell division�1894 – Altmann: first described mitochondria.�1874 – Flemming: described chromosome behaviour during mitosis.�1898 – Golgi: described the Golgi apparatus.

1925 − Gorter & Grendel: described the basic structure of the plasma membrane.

1945 − Porter et al. pioneers in this field of electron microscopy and were the first to identify the endoplasmic reticulum and many elements of the cytoskeleton.

Cell biologist today are still researching the cell.

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References: Mullock BM & Luzio JP, Theory of Organelle Biogenesis: a Historical Perspective (2005). Landes Biosciences, P. 1-13.

Muzzarello P, A Unifying Concept: the History of Cell Theory (1999). Nature Cell Biology, 1, E13-E15.

Exploring the History of Cell Biology:

Timeline

Molecular Biology – A Journey

• Microscopic biology began in 1665
• Robert Hooke (1635-1703) discovered organisms are made up of cells

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• Matthias Schleiden (1804-1881) and Theodor Schwann (1810-1882) further expanded the study of cells in 1830s

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Robert Hooke

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Theodor Schwann

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Matthias Schleiden

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Major events in the history of Molecular Biology 1800 - 1870

• 1865 Gregor Mendel discover the basic rules of heredity of garden pea.
• An individual organism has two alternative heredity units for a given trait (dominant trait vs. recessive trait)

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• 1869 Johann Friedrich Miescher discovered DNA and named it nuclein.

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Mendel: The Father

of Genetics

Johann Miescher

Major events in the history of Molecular Biology 1880 - 1900

• 1881 Edward Zacharias showed chromosomes are composed of nuclein.

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• 1899 Richard Altmann renamed nuclein to nucleic acid.

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• By 1900, chemical structures of all 20 amino acids had been identified

Major events in the history of Molecular Biology 1900-1911

• 1902 - Emil Hermann Fischer wins Nobel prize: showed amino acids are linked and form proteins

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• 1911 – Thomas Hunt Morgan discovers genes on chromosomes are the discrete units of heredity

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• 1911 Pheobus Aaron Theodore Lerene discovers RNA

Emil Fischer

Thomas Morgan

Major events in the history of Molecular Biology 1940 - 1950

• 1941 – George Beadle and Edward Tatum identify that genes make proteins

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• 1950 – Edwin Chargaff find Cytosine complements Guanine and Adenine complements Thymine

George Beadle

Edward Tatum

Edwin Chargaff

Major events in the history of Molecular Biology 1950 - 1952

• 1950s – Mahlon Bush Hoagland first to isolate tRNA

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• 1952 – Alfred Hershey and Martha Chase make genes from DNA

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Mahlon Hoagland

Experiment

Major events in the history of Molecular Biology 1952 - 1960

• 1952-1953 James D. Watson and Francis H. C. Crick deduced the double helical structure of DNA

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• 1956 George Emil Palade showed the site of enzymes manufacturing in the cytoplasm is made on RNA organelles called ribosomes.

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James Watson and

Francis Crick

George Emil Palade

Major events in the history of Molecular Biology 1970

• 1970 Howard Temin and David Baltimore independently isolate the first restriction enzyme

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• This means that: DNA can be cut into reproducible pieces at specific site by restriction enzymes called endonuclease
• The pieces can be linked to bacterial vectors and introduced into bacterial hosts.
• This is called (gene cloning or recombinant DNA technology)

Major events in the history of Molecular Biology 1970- 1977

• 1977 Phillip Sharp and Richard Roberts demonstrated that pre-mRNA is processed by the excision of introns and exons are spliced together.

Phillip Sharp

Richard Roberts

Major events in the history of Molecular Biology 1986 - 1995

• 1986 Leroy Hood: Developed automated sequencing mechanism

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• 1986 Human Genome Initiative announced

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• 1995 Moderate-resolution maps of chromosomes 3, 11, 12, and 22 were published
• These maps provide the locations of “markers” on each chromosome to make locating genes easier

Leroy Hood

Major events in the history of Molecular Biology 1995-1996

• 1995 John Craig Venter: First bacterial genomes sequenced

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• 1995 Automated fluorescent sequencing instruments and robotic operations

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• 1996 First eukaryotic genome-yeast-sequenced

John Craig Venter

Major events in the history of Molecular Biology

• Molecular Biology 1997-1999
• 1999 First human chromosome (number 22) sequenced

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• Molecular Biology 2000-2001
• 2001 International Human

Genome Sequencing published

the first draft of the sequence

of the human human genome

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Major events in the history of Molecular Biology 2003- Present

• April 2003 Human Genome Project Completed
• Mouse genome is sequenced.

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• April 2004 Rat genome sequenced.

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• Next-generation sequencing – genomes being sequenced by the dozen

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Molecular Biology and genetics

• DNA: DeoxyriboNucleic Acid RNA: RiboNucleic Acid
• Bases: Adenine (A) and Guanine (G) = Purines

Cytosine (C) & Thymine (T) = Pyrimidines

• RNA has Uracil instead of Thymine
• Base sugar = Nucleoside (Deoxyribonucleoside & Ribonucleoside)
• Base sugar and phosphate unit = nucleotide . Many form a polynucleotide
• Phosphodiester bonds = link nucleotides for nucleic acid polymer
• Hydrogen bonding between paired bases
• Each chain is complimentary

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Pentose sugar

Ribose

Deoxyribose

Phosphate group

Phosphodiester bond

DNA Packaging

DNA replication

DNA Double Helix

Minor Groove

Major Groove

RNA

• DNA transcribed into mRNA by RNA polymerase
• RNA used as a primer in DNA replication
• Ribosomal RNA
• tRNA aids in translation of mRNA into protein

• Chromatin – DNA bound to histones (DNA packaging proteins)
• DNA and histones are organised into nucleosomes. Eight histones = histone octamer
• These are linked by linker DNA

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1. Double helix is unwound and the base-pairs are separated by DNA helicase.
2. Semi-conservative replication
3. Nucleotides are linked covalently by DNA polymerase
4. Lagging and leading strands
5. DNA ligase for the shorts strands on the lagging strand

Purpose of DNA Extraction

To obtain DNA in a relatively purified form which can be used for further investigations, i.e. PCR, sequencing, etc……

Plasmid purification: alkaline lysis

Alkaline conditions denature DNA

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Neutralize: genomic DNA can’t renature (plasmids CAN because they never fully separate)

Simple DNA extraction

Alkaline cell lysis to break open the cells and organelles

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Alcohol/ethanol precipitation: DNA remaining in the aqueous layer is concentrated by ethanol precipitation.

cell growth

cell harvest and lysis

DNA purification

DNA purification: overview

DNA concentration

A comparison of DNA extraction methods used in research labs as opposed to classroom labs

Research

Lysis: Tissue grinder and use detergent

Precipitation Part I: phenol/chloroform extraction to get rid of proteins

Precipitation Part II: addition of salts to interrupt hydrogen bonding between water and phosphates on the DNA

Precipitation Part III: addition of ethanol to pull DNA out of solution

Wash and resuspend: DNA is washed in ethanol, dried (remove salts and other water soluble impurities ), and resuspended in H20 or TE buffer for long-term storage and stability

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Classroom

Lysis: grind in mortar/pestel and use detergent

Precipitation Part I: NONE (chemical are too dangerous!)

Precipitation Part II: addition of salts to interrupt hydrogen bonding between water and phosphates on the DNA

Precipitation Part III: addition of ethanol to pull DNA out of solution

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LYSIS:�In DNA extraction �this step commonly refers to the breaking �of the cellular membranes (most importantly, the plasma and nuclear membranes)

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• Addition of a detergent in the which breaks down the cell membranes (Sodium Hydroxide)
• Detergents are able to disrupt membranes due to the amphipathic (having both hydrophilic and hydrophobic regions) nature of both cellular membranes and detergent molecules. The detergent molecules are able to pull apart the membranes
• The end result of LYSIS is that the contents of the cells are distributed in solution.

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The cell and nuclear membranes have been broken apart, as well as all of the organelle membranes, such as those around the mitochondria.��So what is left?�

• Proteins
• Carbohydrates (sugars)
• DNA
• Cell debris – cellular membranes.

Filtration

Why?

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• To obtain DNA from cell debris
• Cell debris – too large to go through the filter paper
• DNA, protein, lipids and carbohydrates – small enough to go through the filter paper into the filtrate

PRECIPITATION (In a research lab): �This a series of steps where DNA is separated from the rest of the cellular components�

• In a research lab, the first part of precipitation uses phenol/chloroform to remove the proteins from the DNA
• Phenol denatures proteins and dissolves denatured proteins.
• Chloroform is also a protein denaturant

THIS STEP CANNOT BE PERFORMED IN CLASSROOM LABS!!

• The second part of research lab DNA precipitation is the addition of salts
• The salts interrupt the hydrogen bonds between the water and DNA molecules.

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• The DNA is then precipitated from the protein in a subsequent step with isopropanol or ethanol
• In the presence of cations, ethanol induces a structural change in DNA molecules that causes them to aggregate and precipitate out of solution.
• The DNA is pelleted by spinning with a centrifuge and the supernatant removed

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PRECIPITATION (In a classroom lab): �This a series of steps where DNA is separated from the rest of the cellular components�

• In a classroom lab, DNA precipitation involves the addition of salts
• The salts interrupt the hydrogen bonds between the water and DNA molecules.

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• The DNA is then precipitated from the protein in a subsequent step with isopropanol or ethanol
• In the presence of cations, ethanol induces a structural change in DNA molecules that causes them to aggregate and precipitate out of solution.

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• DNA extracted in a classroom lab is not as “clean” as the DNA extracted in a research lab!

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Similarities

Prokaryotes/Bacteria

Eukaryotes

• Plasma membrane (phospholipid bilayer and regulates permeability)
• Genetic material – DNA
• Cell Wall – except animal cells
• Ribosome - catalyse protein synthesis
• Cytoplasm/cytosol – comprising of water, glucose, proteins and ions.

The cell: https://www.youtube.com/watch?v=URUJD5NEXC8

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Differences

Prokaryotes/Bacteria

Eukaryotes

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• Bacteria
• Size: 1-10μm
• Cell Wall (murein)
• No distinct subcellular organelles
• Circular chromosome – nucleoid
• Often plasmids, RNA and Ribosomes
• Unicellular or multicellular
• Escherichia Coli (E. Coli) is most studied bacterium

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• Plants, animals, fungi and protists (algae and protozoa)
• Size: 10-100μm
• Cell Wall – only plants, fungi and protists (cellulose)
• Well defined subcellular compartments bounded by lipid membranes
• Cytoplasm consists of organelles, ribosomes, cytoskeleton (shape, movement and organises many metabolic functions)
• Cytoskeleton: microtubules made of tublin & microfilaments made of actin.
• Most are multicellular
• Differentiate to specialized tissue/cells

Learning resources from the society for general microbiology

Practical microbiology:

http://www.microbiologyonline.org.uk/media/transfer/doc/sgm_basic_practical_microbiology_2.pdf

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Looking at microbes: http://www.microbiologyonline.org.uk/students/microbe-passports-1#/home

http://www.abcam.com/alpha-smooth-muscle-actin-antibody-1a4-fitc-ab8211.html

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Cytoskeleton – tublin and actin

Exploring organelles

Prokaryotes/Bacteria

Eukaryotes

• Nucleus – Cellular DNA. Transcription & processing of RNA. Nuclear pores within the nuclear membrane.
• Mitochondria – Cellular respiration, the oxidation of nutrients to generate energy in the form of adenosine 5’-triphosphate (ATP). 1-2mm in diameter. 1000-2000 per cell. Smooth outer membrane & Inner folded membrane (cristae). Derived from prokaryotes and retain DNA (circular), RNA and protein machinery.
• Endoplasmic Reticulum (ER) – Cytoplasmic membrane system for lipid biosynthesis and xenobiotic metabolism. Smooth and Rough ER. Rough ER has ribosome attached for protein synthesis.
• Golgi Apparatus – Protein and lipids produced are packaged in the Golgi for final destination.
• Lysosomes – Small membrane-bound organelles & bud off from the Golgi. Consist of degradative enzymes for proteins, nucleic acid, lipids and carbohydrates (macromolecules).
• Centrioles – Regulator of the cell cycle and cytoskeletal organisation.

• Nucleoid – composed of circular double-stranded DNA.
• Plasmid DNA – Short circular DNA and replicates independently of the cell genome.
• Mesosome – Folds of the plasma membrane with associated respiration enzymes. Instead of mitochondria.
• Ribosomes – Smaller, scattered throughout the cytoplasm
• Pilli – protein rods for cell-cell attachment and DNA transfer.
• Flagellum – Motility of many bacteria
• Cell Wall – Rigid and made up of murein (polysaccharide cross-linked by peptide chains). Gram-positive thicker walls compared to Gram-negative. Protection from lysozymes and penicillin.
• Capsule – slime layer of mucilage and helps bacteria form colonies.

Explore molecular biology and genetics:

• What is this model demonstrating?
• Genome video: https://www.youtube.com/watch?v=VJycRYBNtwY
• What Molecular and Genetic words/phrases can you stop in this video? – write them down.

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Molecular Biology

• Cells archive information in the form of DNA, which serves as a master set of instructions for building proteins.
• Protein functions: organelle biosynthesis, structural supports, catalysts of biochemical reactions and aid in the cell's internal and external environments/signalling.
• The length of DNA in each cell is 2 metres long.

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Discuss: What Molecular & Genetic words/phrases can you spot in this video?

Exploring the History of Molecular Biology and Genetics

Timelines to explore:

http://www.bioinformatics.nl/webportal/background/timeline.html

http://www.dnai.org/timeline/

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February 28^th 1952 – The double Helix

This is the date when Watson & Crick (at the University of Cambridge) announced the structure of DNA in the Eagle pub, Cambridge

Molecular Biology and genetics

Transcription & translation

Molecular Biology and genetics

Polymerase Chain Reaction (PCR): DNA amplification

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Denaturation at 94^oC

Primer annealing at 55^oC

Polymerization 72^oC by DNA polymerase

Applications:

• Diagnosis
• Therapy
• Historical studies
• Forensic analysis
• Cloning and sequencing
• Research – genome, genetic variants
• Maybe Dinosaur production in the future – Jurassic world!

Molecular Biology and genetics

Figure 2: The polymerase chain reaction (PCR). Invented in the 1980s

Two key innovations facilitated the use of PCR in the laboratory: the discovery of a DNA polymerase that is stable at the high temperatures used in step 1 of PCR and the development of automated thermal cyclers (machines that bring about the rapid temperature changes necessary for the different steps of PCR).© 2014 Nature Education Adapted from Pierce, Benjamin. Genetics: A Conceptual Approach, 2nd ed. All rights reserved.

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PCR is like a photocopier for DNA

DNA electrophoresis

Applications:

• DNA finger printing for Parentage and Forensics
• Genetic cloning – DNA profiling

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https://www.thenakedscientists.com/podcasts/science-scrapbook/how-does-dna-fingerprinting-work

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Genetic finger printing video:

• Mitosis – Cell division https://www.labtube.tv/video/mesc-cell-mitosis

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• Apoptosis – programmed cell death:

https://www.labtube.tv/video/apoptosis

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• Necrosis - premature death of cells due to injury, infection, cancer, infarction, toxins and inflammation.

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• Migration, differentiation etc…

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Molecular Biology and genetics

Epigenetics:

• Genetic control
• Switch genes on or off and determine which proteins are transcribed:
• DNA methylation
• Histone modifications
• Nucleosome positioning
• All critical for regulating gene expression.
• Our cells all have the same DNA, but our bodies contain many different types of cells: neurons, liver cells, pancreatic cells, inflammatory cells, and others.
• Because certain sets of genes that are "turned on" or expressed, as well as other sets that are "turned off" or inhibited.
• Differential expression.

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Reference: http://www.nature.com/scitable/topicpage/epigenetic-influences-and-disease-895#

Restriction enzymes and DNA modifying enzymes (i.e. genetic editing)

For example, EcoRI restriction enzyme was once a defend mechanism to remove foreign DNA in bacteria, but we have genetically engineered them to be useful tools in cloning – NEB (Don Comb).

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Founded in the mid-1970s as a collective of scientists committed to developing innovative products for the life sciences industry, New England Biolabs is now a recognized world leader in the discovery, development and commercialization of recombinant and native enzymes for genomic research.

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https://www.neb.com/about-neb

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PCR, gene expression, cellular analysis, RNA analysis and epigenetics

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Genetic editing tools:

• Over 3 billion base pairs in a single human’s DNA
• It is difficult to pinpoint what sequences are important in health and disease
• Cas9/CRISPR are like scissors for precise gene targeting

Chromatin / Epigenetics Resources

Protein Acetylation

Histone Methylation

Examples of Crosstalk Between Post-translational Modifications

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MAP Kinase Signaling Resources

MAPK/Erk in Growth and Differentiation

G-Protein-Coupled Receptors Signaling to MAPK/Erk

SAPK/JNK Signaling Cascades

p38 MAPK Signaling Pathways

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Apoptosis Resources

Regulation of Apoptosis

Inhibition of Apoptosis

Death Receptor Signaling

Mitochondrial Control of Apoptosis

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Autophagy Resources

Autophagy Signaling

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PI3K / Akt Signaling Resources

PI3K / Akt Signaling

mTOR Signaling

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Translational Control, Protein Synthesis, and RNA Regulation Resources

Regulation of eIF4E and p70 S6 Kinase

Regulation of eIF2

Overview: Translational Control

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Calcium, cAMP and Lipid Signaling Resources

Protein Kinase C Signaling

Phospholipase Signaling

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Cell Cycle, Checkpoint Control and DNA Damage Resources

G1/S Checkpoint

G2M/DNA Damage Checkpoint

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Cellular Metabolism Resources

Insulin Receptor Signaling

AMPK Signaling

Warburg Effect

Hypoxia Signaling

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Stem Cell Markers, Development and Differentiation

Hippo Signaling Pathway

ESC Pluripotency and Differentiation Pathway

Stem Cell & Lineage Markers

Wnt / β-Catenin Signaling Pathway

Notch Signaling Pathway

Hedgehog Signaling Pathway

TGF-β Signaling Pathway

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Immunology and Inflammation

Jak/Stat: IL-6 Receptor Signaling

NF-κB Signaling

Toll-Like Receptor Signaling

B Cell Receptor Signaling

T-Cell Receptor Signaling

Inflammasome Signaling

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Tyrosine Kinase Resources

ErbB / HER Signaling

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Angiogenesis Resources

Angiogenesis

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Adhesion and Extracellular Matrix Resources

Adherens Junction Dynamics

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Neuroscience Resources

Amyloid Plaque and Neurofibrillary Tangle Formation in Alzheimer's Disease

Dopamine Signaling in Parkinson's Disease

Vesicle Trafficking Presynaptic Signaling

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Cytoskeletal Regulation and Vesicle Trafficking Resources

Regulation of Microtubule Dynamics

Regulation of Actin Dynamics

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Nuclear Receptor Resources

Nuclear Receptor Signaling

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Ubiquitin and Ubiquitin-Like Protein Resources

Ubiquitin / Proteasome

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Protein Folding

Organelle Markers

- See more at: https://www.cellsignal.com/common/content/content.jsp?id=science-pathways#sthash.gyZVTqJ7.dpuf

Cell signalling pathways

Explanation to cell and molecular biology research

Organelle Diseases:

Mitochondrial diseases

• Dysfunctional mitochondria
• Mutations in mitochondrial DNA and nuclear DNA
• 1 in 200 children in the UK
• Mitochondrial DNA disease is passed down from mother to child
• Currently no cure
• Unlikely to survive childhood
• The Wellcome Trust Centre for Mitochondrial Research at Newcastle University has developed a treatment known as mitochondrial donation (http://www.wellcome.ac.uk/About-us/Policy/Spotlight-issues/Mitochondrial-diseases/)

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Epigenetics related diseases/disorders

• Cancer
• Mental Retardation
• Obesity
• Immunodeficiency

(http://www.nature.com/scitable/topicpage/epigenetic-influences-and-disease-895)

(http://www.ncbi.nlm.nih.gov/pubmed/20944598)

Cancer

• Epigenetics
• DNA mutations
• Inherited genes
• Cell cycle
• Hormones and growth factors – proliferation, differentiation and apoptosis of cells

http://www.nature.com/scitable/search-scitable?criteria=cancer

Biotechnology and Biopharmaceutical

• Gene and Cell Therapy
• Antibodies for therapy and diagnostics
• Gene engineering – e.g Insulin, cloning
• Stem cells – iPSC
• Personalised medicine
• Diagnosis

Covered in next lecture.

Metabolic Diseases – for example Diabetes

(http://www.nlm.nih.gov/medlineplus/metabolicdisorders.html)

Cell cycle related diseases/disorders:

• Cancer
• Cardiovascular diseases
• Infection
• Inflammation
• Neurodegenerative

(http://www.ncbi.nlm.nih.gov/pubmed/20964732)

Cell signalling diseases/disorders– for example CVD, tissue repair, neurological

Publicity

• Nature (http://www.nature.com/)
• New Scientist (http://www.newscientist.com/)
• Naked Scientist (http://www.thenakedscientists.com/)
• LabTube (https://www.labtube.tv/)

Search engines for journals

• Pubmed (http://www.ncbi.nlm.nih.gov/pubmed)
• Google Scholar (https://scholar.google.co.uk/)

Biotechology websites

• Life Technologies

(https://www.lifetechnologies.com/uk/en/home.html)

• Abcam

http://www.abcam.com/

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• New England Biolabs (NEB) (http://www.neb.uk.com/)
• Illumina (http://www.illumina.com/)
• Horizon (http://www.horizondiscovery.com/ )

Company websites

• GSK (http://uk.gsk.com/)
• Medimmune (https://www.medimmune.com/)
• Babraham Research park (http://www.babraham.co.uk/about/about.html)
• Pfizer (http://www.pfizer.co.uk/)
• OxfordBiomedica (http://www.oxfordbiomedica.co.uk/)

How to research the cellular and molecular universes yourself:

Institute websites

• Wellcome Trust Sanger Institute (https://www.sanger.ac.uk/)
• Ensembl (http://www.ensembl.org/index.html) Genome databases for vertebrates and other eukaryotic species.
• Human Genome (https://www.genome.gov/11006943)
• British Society of Gene and Cell Therapy (http://www.bsgct.org/en/)
• European Society of Gene and Cell Therapy(http://www.esgct.eu/)
• American Society of Gene and Cell Therapy (http://www.asgct.org/)

‘UNIVERSITY CHALLENGE’ QUIZ

• Form groups – four teams required
• Are you ready for the cell and molecule biology quiz?!

References

• Bianconi E. et al. An estimation of the number of cells in the human body (2013). Ann Hum Biol; 40 (6): 463-71.
• W. R. Pickering. Oxford Revision guides, AS & A level Human Biology through diagrams.
• Turner P. et al. Bios Instant Notes, Molecular Biology, 3^rd edition.
• All images are from Clip Art on MicroSoft Word

Question two: Name these organelles (1 point for each organelle)?

Simple DNA extraction

Alkaline cell lysis to break open the cells and organelles

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Alcohol/ethanol precipitation: DNA remaining in the aqueous layer is concentrated by ethanol precipitation.