Application of Site-directed Mutagenesis to RNA and DNA Genomes (original) (raw)

1. H. Weber,
2. T. Taniguchi,
3. W. Müller,
4. F. Meyer, and
5. C. Weissmann
6. Institut für Molekularbiologie I, Universität Zürich, 8093 Zürich, Switzerland

Excerpt

The elucidation of the relationship between the structure and function of biological macromolecules is greatly aided by the availability of appropriate mutants. The classical genetic approach depends on mutations that arise at random either spontaneously or as a consequence of mutagenic agents. Mutants are usually selected because of some desired phenotype or striking property. Difficulties may arise when the role of noncoding, functionally undefined segments of a genome are to be studied, since one does not know what property to screen or select for. Moreover, in haploid organisms, unconditionally lethal mutations cannot be picked up at all. The problems are compounded in the case of higher organisms, where recessive mutations are frequently only recognizable when homozygous. In addition, the genome of higher eukaryotes is so large that the probability of finding a mutation in a particular DNA region without a potent selection system is quite low.

The advent of hybrid...