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An Introduction to Molecular Biotechnology


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A P Posttranslational modification of proteins, modification of sugar chains Lysosome A Harbors hydrolytic enzymes, degrades organelles and macromolecules, macrophages eat invading microbes Vacuole P Sequestration of storage proteins, defense and signal molecules, contains hydrolytic enzymes, degrades organelles and macromolecules Mitochondrium A P Organelle derived from endosymbiotic bacteria; contains circular DNA, own ribosomes; enzymes of citric acid cycle, β‐oxidation, and respiratory chain (ATP generation) Chloroplast P Organelle derived from endosymbiotic bacteria; contains circular DNA, own ribosomes; chlorophyll and proteins of photosynthesis, enzymes of CO2 fixation and glucose formation (Calvin cycle) Peroxisome A P Contains enzymes that generate and degrade H2O2 Cytoplasm A P Harbors all compartments, organelles, and the cytoskeleton of a cell; many enzymatic pathways (e.g. glycolysis) occur in the cytoplasm

      A, animal; P, plant.

      The most important biochemical and cell biological characters of Archaea, Bacteria, and Eukarya are summarized in Table 1.1.

Image described by caption.

      Eukaryotic cells are characterized by compartments that are enclosed by biomembranes (Table 1.2). As a result of these compartments, the multitude of metabolic reactions can run in a cell at the same time.

      In the following discussion on the shared characteristics of all cells, the diverse differences that appear in multicellular organisms should not be forgotten. The human body has more than 200 different cell types, which show diverse structures and compositions. These differences must be understood in detail if cell‐specific disorders, such as cancer, are to be understood and consequently treated. Modern technology with Next‐Generation Sequencing (NGS) allows a study of single cells at a genomic and transcriptomic level.

      Before a detailed discussion of cellular structures and their functions (see Chapters 35), a short summary of the biochemical basics of cellular and molecular biology is given in Chapter 2.



Problem Technique/instrument Remarks
Structure elucidation of proteins Protein isolation, column chromatography (gel filtration, ion exchange, affinity) Chapter 7
Gel electrophoresis Chapter 7
Protein–protein interactions (FRET, two hybrid systems, FRAP) Chapters 19 and 23
Crystallization
X‐ray diffraction
NMR
Cryoelectron microscopy
Mass spectrometry Chapter 8
Protein sequencing
DNA PCR and quantitative PCR (qPCR) Chapter 13
DNA/RNA isolation Chapter 9
DNA hybridization Chapter 11
Sanger sequencing Chapter 14
Restriction enzymes