Gene Mapping 793
never come into actual physical contact. The genetic
data can nevertheless provide evidence for involve-
ment in the same pathway or process.
See also: Alleles; Dominance; Epistasis; Recessive
Inheritance; Suppression; Suppressor Mutations
Gene Library
I Schildkraut
Copyright ß 2001 Academic Press
doi: 10.1006/rwgn.2001.0512
A gene library is a collection of single cells (usually
bacteria), each of which has received a single segment
of DNA usually carried by a plasmid, bacteriophage,
or viral vector, the DNA segments having been
derived from genomic DNA or cDNA.
See also: Genomic Library
Gene Mapping
J R S Fincham
Copyright ß 2001 Academic Press
doi: 10.1006/rwgn.2001.0513
The Detection of Recombination within
Genes
In the sense intended here, gene mapping means the
ordering of sites within genes. In the earlier history of
genetics, the gene was considered to be a single indi-
visible unit of mutation and recombination, but from
the early 1950s onwards it became apparent that dif-
ferent mutations in the same gene were nearly always
at different sites and able to recombine to yield wild-
type and, where they were looked for, doubly mutant
genes. In eukaryotic organisms (e.g., fungi and Dros-
ophila) the recombinants were generated in meiosis
following a sexual cross between mutants. In bacteria,
recombination occurred in the course of conjugation,
transduction, or transformation, as the donor genomic
fragment was integrated into the whole genome of the
recipient cell. In bacteriophage it occurred during
mixed infection of bacterial cells.
The frequency of recombination within genes is
low compared with that between genes. In the budding
yeast Saccharomyces cerevisiae it may amount to a few
percent of the meiotic products, but in most other
eukaryotes favored by geneticists it is very much
lower. Consequently, the use of recombination for
mapping within genes depends on some method for
selecting wild-type recombinants from a large excess
of nonrecombinant mutants. This is straightforward
when the mutants have a growth handicap not shared
by the wild-type, as when they have special nutritional
requirements (i.e., are auxotrophs) or are sensitive to
higher temperatures, or (in bacteriophage genetics)
unable to grow in a particular host.
Mapping by Recombination Frequency
It is a general principle of linkage mapping that the
frequency of recombination between sites of mutation
increases with their distance apart. But the use of
recombination frequency for mapping within genes
is complicated by the fact that the sites being recom-
bined are close to the recombination event, which is a
complex process probably always involving local non-
reciprocality. In fungi and Drosophila, which are the
eukaryotic organisms most studied in this regard,
much or most recombination between mutant sites
in the same gene (usually detected as production of
wild-types from intermutant crosses) is due to the
nonreciprocal conversion of one mutant site to wild-
type. The contribution of reciprocal crossing-over is
greater when the mutant sites are relatively widely
spaced.
However, even if most recombination within genes
is due to conversion and not to crossing-over, we still
expect, and generally find, a strong correlation between
recombination frequency and distance. The reason is
that conversion involves tracts of DNA rather than
single base pairs, and recombination between two sites
by conversion will occur only when the conversion
tract covers one but not the other, and this will ob-
viously be less likely the closer the spacing of the sites.
In practice, recombination frequency is a good gen-
eral guide to gene mapping but sometimes gives
ambiguous results. One source of confusion is that the
nature of the mutational site may strongly influence its
probability of conversion (see Gene Conversion and
Mismatch Repair (Long/Short Patch)). This is called a
marker effect.
The Use of Flanking Markers
To the extent that recombination between mutant sites
within a gene is due to reciprocal crossing-over, it
should result in recombination of genetic markers
placed on either side of the gene. Thus, wild-type
recombinants should be associated with one new flank-
ing marker combination, and double-mutant recom-
binants (if they are recoverable) with the reciprocal