An Introduction to Molecular Biotechnology

Fundamentals, Methods and Applications

Michael Wink, Michael Wink

• Publisher: Wiley-VCH
• Year: 2020
• ISBN:
  9783527812882
• Edition: 3
• Topic: Biological Sciences
• Subtopic: Biotechnology
• Index: Biological Sciences

Completely updated in line with the rapid progress made in the field, this new edition of the highly-praised textbook addresses powerful new methods and concepts in biotechnology, such as genome editing, reprogrammed stem cells, and personalized medicine.
An introduction to the fundamentals in molecular and cell biology is followed by a description of standard techniques, including purification and analysis of biomolecules, cloning techniques, gene expression systems, genome editing methods, labeling of proteins and in situ-techniques, standard and high resolution microscopy. The third part focuses on key areas in research and application, ranging from functional genomics, proteomics and bioinformatics to drug targeting, recombinant antibodies and systems biology. The final part looks at the biotechnology industry, explaining intellectual property issues, legal frameworks for pharmaceutical products and the interplay between start-up and larger companies. The contents are beautifully illustrated throughout, with hundreds of full color diagrams and photographs.
Provides students and professionals in life sciences, pharmacy and biochemistry with everything they need to know about molecular biotechnology.

Part I
Fundamentals of Cellular and Molecular Biology

1
The Cell as the Basic Unit of Life

Michael Wink

Heidelberg University, Institute of Pharmacy and Molecular Biotechnology (IPMB), Im Neuenheimer Feld 329, 69120 Heidelberg, Germany

The base unit of life is the cell. Cells constitute the base element of all prokaryotic cells (cells without a cell nucleus, e.g. Bacteria and Archaea) and eukaryotic cells (or Eukarya) (cells possessing a nucleus, e.g. protozoa, fungi, plants, and animals). Cells are small, membrane‐bound units with a diameter of 1–20 μm and are filled with concentrated aqueous solutions. Cells are not created de novo, but possess the ability to copy themselves, meaning that they emerge from the division of a previous cell. This means that all cells, since the beginning of life (around 4 billion years ago), are connected with each other in a continuous lineage. In 1885, the famous cell biologist Rudolf Virchow conceived the law of omnis cellula e cellula (all cells arise from cells), which is still valid today.

The structure and composition of all cells are very similar due to their shared evolution and phylogeny (Figure 1.1). We see an astonishing constancy in fundamental structures and mechanisms. Owing to this, it is possible to limit the discussion of the general characteristics of a cell to a few basic types (Figure 1.2):

• Bacterial cells
• Plant cells
• Animal cells

Figure 1.1 Tree of life – phylogeny of life domains.

Figure 1.2 Schematic structure of prokaryotic and eukaryotic cells. (a) Bacterial cell, (b) plant mesophyll cell, and (c) animal cell.

Nucleotide sequences from 16S rRNA, amino acid sequences of cytoskeleton proteins, and characteristics of the cell structure were used to reconstruct this phylogenetic tree. Prokaryotes are divided into Bacteria and Archaea. Archaea form a sister group with eukaryotes; they share important characteristics (Tables 1.1 and 1.2). Many monophyletic groups can be recognized within the eukaryotes (diplomonads/trichomonads, Euglenozoa, Alveolata, Stramenopilata [heterokonts], red algae and green algae/plants, fungi and animals; see Tables 6.3–6.5 for details).

Table 1.1 Comparison of important biochemical and molecular characteristics of the three domains of life.

Character	Prokaryotes		Eukaryotes
	Archaea	Bacteria
Organization	Unicellular	Unicellular	Unicellular or multicellular
Cytology
Internal membranes	Rare	Rare	Always (Table 1.2)
Compartments	Only cytoplasm	Only cytoplasm	Several (Table 1.2)
Organelles	No	No	Mitochondria; plastids
Ribosomes	70S	70S	80S (mt, cp: 70S)
Membrane lipids	Ether lipids	Ester lipids, hopanoids	Ester lipids, sterols
Cell wall	Pseudopeptidoglycan, polysaccharides, glycoproteins	Murein (peptidoglycan), polysaccharides, proteins	PL: polysaccharides, cellulose F: chitin A: no
Cytoskeleton	FtsZ and MreB protein	FtsZ and MreB protein	Tubulin, actin, intermediary filaments
Cell division	Binary fission	Binary fission	Mitosis
Genetics
Nuclear structure	Nucleoid	Nucleoid	Membrane‐enclosed nucleus with chromosomes
Recombination	Similar to conjugation	Co...