Process for in vitro creation of recombinant polynucleotide sequences by oriented ligation
First Claim
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1. A ligase-mediated method of recombination, comprising:
- providing oligonucleotide fragments derived from each of at least two heterologous polynucleotide sequences of a polynucleotide bank;
hybridizing the fragments to an assembly matrix so that the hybridized fragments are oriented for ligation with each other; and
ligating the hybridized fragments having immediately adjacent ends with a ligase to form random recombinant polynucleotide sequences.
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Abstract
The object of this invention is a process for the creation of at least one recombinant polynucleotide sequence comprising a step of oriented ligation of fragments derived from a bank of at least two polynucleotide sequences, and optionally cloning the recombinant polynucleotide sequences, and the selection of polynucleotide sequences offering advantageous characteristics compared to one or several reference sequences.
94 Citations
44 Claims
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1. A ligase-mediated method of recombination, comprising:
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providing oligonucleotide fragments derived from each of at least two heterologous polynucleotide sequences of a polynucleotide bank; hybridizing the fragments to an assembly matrix so that the hybridized fragments are oriented for ligation with each other; and ligating the hybridized fragments having immediately adjacent ends with a ligase to form random recombinant polynucleotide sequences. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 44)
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43. A ligase-mediated method of recombination, comprising:
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hybridizing oligonucleotide fragments derived from each of at least two heterologous polynucleotide sequences to an assembly matrix so that the hybridized fragments are oriented for ligation with each other; and ligating the hybridized fragments having immediately adjacent ends with a ligase to form random recombinant polynucleotide sequences.
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Specification