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What's new?: From gene to phenotype in Drosophila and other organisms
Author(s) -
Kaiser Kim
Publication year - 1990
Publication title -
bioessays
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.175
H-Index - 184
eISSN - 1521-1878
pISSN - 0265-9247
DOI - 10.1002/bies.950120609
Subject(s) - transposable element , biology , genetics , transposon mutagenesis , gene , locus (genetics) , mutagenesis , population , sleeping beauty transposon system , dna , mutant , phenotype , polymerase chain reaction , demography , sociology
Abstract The growing number of cloned eukaryotic genes lacking a defined or proven biological function poses a major challenge in ‘reverse genetics’. A method is described here that permits efficient screening for new lesions in, or close to, genes corresponding to cloned DNA sequences of interest. The technique involves transposon mutagenesis, followed by screening of DNA isolated from a population of mutagenised individuals (or their progeny) for evidence that the population contains at least one individual in which transposon insertion has occurred at the target locus. Detection of rare individuals within the population is facilitated by the use of the polymerase chain reaction (PCR). Once recognised, specific individuals (or their progeny) are isolated from the population by a process of sib‐selection. In cases where insertion of the transposon has occurred close to, but not within, the target locus, secondary events involving imprecise excision of the transposon will nonetheless allow the isolation of mutant individuals. Though the method was developed specifically for the transposon‐mutagenesis of Drosophila , extensions to other organisms and to other mutagenic strategies are feasible and some of the possibilities are discussed.