Interference No. 105,188
Short v. Punnonnen
1. Replicating a clone containing a nucleotide
sequence under conditions that allow mutations to occur.
2. Replicating a second clone containing a second
nucleotide sequence under conditions that allow mutations to
5 occur.
3. Randomly combining and co-expressing the two
mutated populations of nucleotide sequences.
4. Screening clones containing combinations of the
mutated nucleotide sequences for phenotypes that were not
10 present in either parent clone.
Alternatively, the methods combine the following
elements:
15 1. Replicating at least portions of two nucleotide
sequences contained within a single clone under conditions
that allow mutations to occur in either nucleotide sequence.
2. Allowing recombination events between the two
nucleotide sequence populations to reassociate mutant
20 nucleotide sequences to form new pairs of the two sequences
that were not paired in the original mutated, replicated
population.
3. Screening clones containing combinations of
nucleotide sequences for phenotypes that were not present in
25 the parent clone or in the mutant replicas of the parent
clone.
For example, assume a parent clone containing two
nucleotide sequences A and B is replicated under mutating
30 conditions such that variant clones are formed:
Parent: A/B
Variant 1: A1/B
Variant 2: A/B1
35 Variant 3: A2/B1
Variant 4: A/B2
Variant 5: A3/B
However, within this mutated population, the combinations
40 A1/B2, A2/B, A2/B2, A3/B1, and A3/B2, do not occur. If the
mutant population (including non-mutated parent clones) is
allowed to recombine sequences A and B and their variants,
then combinations such as A1/B2, A2/B etc. can be created.
Such new combinations may express a desired phenotype that
45 was not present in the parental or variant population.
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Last modified: November 3, 2007