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Summary: Recent in vitro recombination or "DNA shuf-
fling" experiments have demonstrated that
exchanging fragments of closely homologous
genes provides an efficient way to generate
proteins with new traits1,2. The resulting mole-
cules are very different, at least in sequence,
from those that might be obtained by more
local searches of protein space, for example by
random mutagenesis. The DNA shuffling
method, which relies on homologous recom-
bination during the PCR reassembly of gene
fragments from multiple parents, generates
crossovers at points of high sequence identity
(Fig. 1A). Would crossovers between regions
of low or no homology also be useful for mole-
cular breeding of highly similar genes, or to
shuffle more distantly related and even unre-
lated sequences? Would this approach generate
a more efficient search of sequence space and,
practically, more rapid adoption of novel pro-
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