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Chapter_08_Solutions - Chapter 8 Manipulating Proteins DNA...

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ISOLATING CELLS AND GROWING THEM IN CULTURE DEFINITIONS 8–1 Monoclonal antibody 8–2 Hybridoma TRUE/FALSE 8–3 False. A monoclonal antibody recognizes a specific antigenic site, but this does not necessarily mean that it will bind only to one specific protein. There are two complicating factors. First, antigenic sites that are similar, but not identical, can bind to the same antibody with different affinities. If too much antibody is used in an assay, the antibody may bind to one protein with high affinity and to others with low affinity. Second, it is not uncommon for different proteins to have the same antigenic site; that is, the same clus- ter of five or six amino acid side chains on their surfaces. This is especially true of members of protein families, which have similar amino acid sequences, and are often identical in functionally conserved regions. 8–4 True. The two techniques differ in what is done with the very early embryo. For reproductive cloning the embryo is placed in a foster mother, who (very rarely) will give birth to an individual who is a clone of the individual that donated the original somatic cell nucleus. For therapeutic cloning the embryo is used as a source of embryonic stem (ES) cells, with the aim of gen- erating various cell types that can be used for tissue repair. These so-called personalized ES cells will be genetically identical to the original nuclear donor. In principle, tissues generated from such cells could be used thera- peutically in the adult from whom the original nucleus came, without fear of tissue rejection. THOUGHT PROBLEMS 8–5 Cells in a tissue are bound together by protein-mediated attachments to one another and to an extracellular matrix containing collagen. Treatment with trypsin, collagenase, and EDTA disrupts these attachments. Trypsin is a pro- tease that will cleave most proteins, but generally only those portions of a native protein that are unstructured. The triple-helical structure of collagen, for example, is a poor substrate for trypsin. Collagenase, which is a protease specific for collagen, digests a principal component of the extracellular matrix. EDTA chelates Ca 2+ , which is required for the cell-surface proteins known as cadherins to bind to one another to link cells together. Removal of Ca 2+ prevents this binding and thereby loosens cell–cell attachments. A183 In This Chapter ISOLATING CELLS A183 AND GROWING THEM IN CULTURE PURIFYING PROTEINS A185 ANALYZING PROTEINS A189 ANALYZING AND A195 MANIPULATING DNA STUDYING GENE A208 EXPRESSION AND FUNCTION Chapter 8 8 Manipulating Proteins, DNA, and RNA
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A184 Chapter 8: Manipulating Proteins, DNA, and RNA The treatment doesn’t kill the cells because all the damage occurs to extra- cellular components, which the cells can replace. So long as the plasma membrane isn’t breached, the cells will survive.
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Chapter_08_Solutions - Chapter 8 Manipulating Proteins DNA...

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