Titlebook: Genetic Engineering Cloning DNA; David M. Glover Book 1980 D. M. Glover 1980 DNA.Expression.chromosome.engine.eukaryote.genetic engineerin

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https://doi.org/10.1007/978-3-642-91017-3Bacteriophage λ has about 50 genes in its 49Kb genome, only about half of which are essential for its lytic growth. Before describing λ vectors in which this non-essential DNA can be replaced by foreign DNA, I will give a simplified description of the life cycle of the bacteriophage. More comprehensive reviews can be found elsewhere [1,2].

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,Bacteriophage λ vectors,Bacteriophage λ has about 50 genes in its 49Kb genome, only about half of which are essential for its lytic growth. Before describing λ vectors in which this non-essential DNA can be replaced by foreign DNA, I will give a simplified description of the life cycle of the bacteriophage. More comprehensive reviews can be found elsewhere [1,2].

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In-Situ

The enzymology of , DNA recombination,fic sequences in DNA and then cleave both strands of the duplex. These enzymes have been found in many prokaryotes and are likely to be responsible for the degradation of ‘alien’ DNA molecules, the indigenous DNA being protected from degradation by a modification enzyme, usually a methylase. Restric

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Plasmid vectors,ndonuclease cleavage sites in regions non-essential for DNA replication or for the marker function. Most vectors are non-conjugative plasmids and so do not bring about the transfer of DNA from one cell to another. This is a desirable property with respect to the containment of any potentially hazard

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Expression of cloned DNAs in ,nslational machinery of the host cell. The bacterium . is a natural choice for such studies because of our detailed knowledge of the molecular biology of its gene expression. This chapter will examine the suitability of the most commonly used cloning vehicles as systems for expressing foreign genes.

PATRI

Methods for the physical characterisation of cloned segments of chromosomal DNA from higher eukaryoouse, for example, contains approximately 3 × 10.Kb of DNA, and so the purification and characterisation of a gene such as that for β globin was impossible before the advent of such techniques. The cloning technology not only allows the purification of the gene but also its amplification during prop

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