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The single sentence definition of biotechnology: “the application of science and technology to living organisms, as well as parts, products and models thereof, to alter living or non-living materials for the production of knowledge, goods and services.” OECD list-based definition of biotechnology techniques: DNA/RNA: Genomics, pharmacogenomics, gene probes, genetic engineering, DNA/RNA sequencing/synthesis/amplification, gene expression profiling, and use of antisense technology. Proteins and other molecules: Sequencing/synthesis/engineering of proteins and peptides (including large molecule hormones); improved delivery methods for large molecule drugs; proteomics, protein isolation and purification, signaling, identification of cell receptors. Cell and tissue culture and engineering: Cell/tissue culture, tissue engineering (including tissue scaffolds and biomedical engineering), cellular fusion, vaccine/immune stimulants, embryo manipulation. Process biotechnology techniques: Fermentation using bioreactors, bioprocessing, bioleaching, biopulping, biobleaching, biodesulphurisation, bioremediation, biofiltration and phytoremediation. Gene and RNA vectors: Gene therapy, viral vectors. Bioinformatics: Construction of databases on genomes, protein sequences; modelling complex biological processes, including systems biology. Nanobiotechnology: Applies the tools and processes of nano/microfabrication to build devices for studying biosystems and applications in drug delivery, diagnostics etc. Source: OECD (2005), A Framework for Biotechnology Statistics, OECD, Paris. See also this link: http://www.oecd.org/dataoecd/4/23/42833898.pdf Biotechnology is mostly about cells or parts of them, but entire organisms may also in some cases be seen as parts of biotechnology. In either case it is typical that one looks at cells or organisms from a chemical point of view, trying to explain observations in molecular terms. This often leads into an enormous complexity, which has resulted in a focus on multidiciplinarity (biology, chemistry, bioinformatics, physics, mathematics) Biotecnology originates from classical biology, which mainly has been a descriptive field, but now there is room for everyone! Tree of life PROKARYOTES Bacteria EUKARYOTES Archaea Entamoebae Slime molds Green nonsulfur bacteria Mitochondrion Proteobacteria Chloroplast Flavobacteria Cyanobacteria Grampositive bacteria Eukarya Euryarchaeota Methanosarcina MethanoExtreme Crenarchaeota bacterium halophiles Thermoproteus Methanococcus Pyrodictium Thermoplasma Animals Fungi Plants Ciliates Thermococcus Marine Crenarchaeota Flagellates Pyrolobus Methanopyrus Trichomonads Thermotoga Thermodesulfobacterium Microsporidia Aquifex Universal ancestor Diplomonads (Giardia) Cytoplasm Cell wall Nucleoid Ribosomes Plasmid Cytoplasmic membrane Prokaryote Cytoplasmic membrane Endoplasmic reticulum Ribosomes Nucleus Nucleolus Nuclear membrane Cytoplasm Eukaryote Golgi Mitochondrion Chloroplast The structures of animal and plant cells, which both are eukaryotic organisms A short outline of eukaryotic organelle functions: Cell wall: A stiff structure outside of the cytoplasmic membrane in plant cells (and bacteria). Ensures a specific form of the cell and also contributes to stiffness of tissues (like wood). Centriole: An organelle involved in separation of chromosome copies in mitosis and other ”motor” activities. Contains microtubules (fibrous motor protein complexes), also found elsewhere in cells. Chloroplast: An organelle that contains DNA and is responsible for photosynthesis in plant cells. Chromatin: Complex of DNA and portiens (such as histones) in the nucleus. Chromosome: A particularly condensed form of the DNA (and associated proteins). Endoplasmatic reticulum (ER): Network of interconnected membranestructures in the cytoplasm. Rough ER is associated with ribiosomes and functions in synthesis of secretory and membrane proteins. Smooth ER: lacks ribosomes and is involved in lipid biosynthesis. Flagellum: Protein complex that can be used for cell movement. Golgi complex/Golgi apparatus: Stacks of mebraneous structures that function in processing and sorting of lipids and proteins to be sent to specific cellular compartments or for secretion. Lysosome: A membrane, bounded ”bag” with an internal low pH and which can ingest and digest particles entering the cell. Microfilament: Protein fibers acting as motors, for example in muscle contraction. Mitochondrion: Organelle surrounded by a membrane similar to in bacteria. Contains a small DNAmolecule and is the site of oxidative phosphorylation, leading to ATP formation. Nucleolus: Structure in the nucleus responsible for rRNA synthesis and ribosome subunit assembly. Nucleus: A membrane-bounded organelle containing the chromosomes Peroxisome: Involved in degradation of fatty acids and amino acids. Plasma membrane. The membrane that represents the outer boundary of the cell. Plasmodesmata: Channles for transport out of or between cells in plants. Ribosome: A particle consisting of over 50 different proteins and rRNA and that represents the machinery for translation. Tonoplast: The mebrane that surrounds the vacuole. Vacuole: A ”bag”/vesicle inside the cytoplasm involved in intracellular secretion, storage and digestion. A cut through an onion: Organisms are collections of cells that cooperate. It is therefore critical to understand how a single cell functions. The structures of bacteria are much simpler than those of eukaryotes. Bacteria lack organelles, and there are also many other fundamental differences in the entire cellular machinery, molecularly and functionally. Thus, from a molecular point of view humans are much more similar to bananas than bananas are to bacteria. The mitochondria in eukaryotes are probably ”old” bacteria. All organisms (except some viruses, which are normally not called organisms) still share in common that their hereditary material is DNA. DNA (nucleoid) Cell wall Cytoplasmic membrane The typical appearance of bacteria as seen in a transmission electron microsocpe (a) or in a scanning electron microscope (c) Two main types of bacteria: G+ and G- Gram–negative Gram–positive Peptidoglycan Peptidoglycan Cytoplasm Cytoplasm Cytoplasmic membrane Periplasm Membrane Outer membrane (Iipopolysaccharide and protein) Outer membrane Peptidoglycan Cytoplasmic membrane Cytoplasmic membrane Peptidoglycan Archae are neither bacteria nor eukaryotes, but belong to the prokaryotes, see tree of life Archae can form numerous “bisarre” structures Viruses can attack all types of organisms, from bacteria to plants, animals and humans. Any given virus typically attacks only one or a few closely related species. All viruses share in common that they depend on a cell to propagate themselves. Unlike all cells, viruses may have RNA as their hereditary material FIGURE 1.24 Examples of Different Viruses Viruses come in a variety of shapes and sizes that determine whether the entire virus or only its genome enters the host cells. Biotechnology by Clark and Pazdernik Copyright © 2012 by Academic Press. All rights reserved. 13 Viruses may contain many different forms of their herediatry material (DNA or RNA), and it can consist of a single chromosome or several, and the DNA or RNA may be single or double-stranded. In cells the hereditary material is always double-stranded DNA. Nonenveloped Enveloped ssDNA Parvovirus Nonenveloped Enveloped all ssRNA partially dsDNA Hepadnavirus Rhabdovirus ssRNA dsDNA Papovavirus dsDNA Picornavirus Togavirus Orthomyxovirus dsDNA Poxvirus Adenovirus dsRNA dsDNA Bunyavirus Coronavirus Arenavirus Retrovirus Reovirus dsDNA Herpesvirus Paramyxovirus Iridovirus DNA viruses RNA viruses T4-virus, is a bacteriophage, which means it attacks bacteria Influenza-virus Infection of a bacterium with bacteriophage T4 Tail fibers Tail pins Outer membrane Tail lysozyme Peptidoglycan Cytoplasmic membrane Cytoplasm T4 genome Living organisms are built from cells, and from a scientific point of view they may be compared to machines (cars?) Car Can make copies of themselves? Need energy to function? The energy originates from the same sources? Need energy to maintain themselves? Functions according to the laws of nature? Wears out over time, even if energy supply is good? Can repair itself? Cell The chemical components of cells: Macromolecules: Nucleic acids (DNA and RNA) Proteins Lipids Polysaccharides Lipopolysaccharides (a combination of lipid and polysaccharide) ”Small” molecules (including metabolites) and elements Water Carbohydrates (also building blocks of polysaccharides) Amino acids (also building blocks of proteins) Purins/pyrimidines (also parts DNA and RNA) Nukleosides and nukleotides (also parts of DNA and RNA, contain purins and pyrimidins) Other organic metabolites Phosphate Sulfate Ammonium Metals/ions