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Delitto PerfettoTargeted Mutagenesis in Yeast with Oligonucleotides

2003; Linguagem: Inglês

10.1007/978-1-4615-0073-5_9

0887-8307

Francesca Storici, Michael A. Resnick,

Chromosomal and Genetic Variations

Now that the genomes of many organisms have been completely sequenced, opportunities to examine functions of specific genes and to investigate genome organization have been greatly increased. Gene targeting and site-directed mutagenesis have been instrumental in dissecting genetic pathways, studying gene regulation and analyzing protein structure-function relationships. Stable modifications of chromosomal DNA are preferable to the addition of unstable vectors containing a modified version of a gene or DNA of interest. Moreoverin situgene engineering allows physiological expression of the targeted DNA sequence, since it remains under control of natural regulatory regions. Approaches directed at modifying genomes should provide for rapid, accurate and multiple changes without the introduction of unwanted or heterologous DNA, such as vector DNA or sequences from selectable markers. In fact, one of the concerns in genetic engineering is the possibility that foreign DNA could be expressed in the genome of the genetically altered organism.