Incrementally truncated nucleic acids and methods of making same
First Claim
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1. A method of making a plurality of incrementally truncated hybrid nucleic acids comprising the steps of:
- a) serially removing nucleotides from one or both termini of a first and a second parent nucleic acid to form truncated first and second parent nucleic acids whose length decreases incrementally over time;
b) stopping said serial nucleotide removal at a plurality of different times to form a plurality of incrementally truncated first and second parent nucleic acids;
c) linking separate incrementally truncated first parent nucleic acids to separate incrementally truncated second parent nucleic acids to form a plurality of incrementally truncated hybrid nucleic acids.
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Abstract
A series of methods that utilize the incremental truncation of nucleic acids are described to create a plurality of modified nucleic acids and hybrid polypeptides. A plurality of substantially all possible single base-pair deletions of a given nucleic acid sequence is created. A method of making shuffled incremental truncated nucleic acids, which is independent of nucleic acid sequence homology, is also described. These methods can be used in protein engineering, protein folding, protein evolution, and the chemical synthesis of novel hybrid proteins and polypeptides.
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9 Claims
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1. A method of making a plurality of incrementally truncated hybrid nucleic acids comprising the steps of:
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a) serially removing nucleotides from one or both termini of a first and a second parent nucleic acid to form truncated first and second parent nucleic acids whose length decreases incrementally over time; b) stopping said serial nucleotide removal at a plurality of different times to form a plurality of incrementally truncated first and second parent nucleic acids; c) linking separate incrementally truncated first parent nucleic acids to separate incrementally truncated second parent nucleic acids to form a plurality of incrementally truncated hybrid nucleic acids. - View Dependent Claims (2, 3)
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4. A method of making a plurality of first variant incrementally truncated hybrid nucleic acids comprising the steps of:
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a) providing a first and second parent nucleic acid; b) serially removing nucleotides from one or both termini of said first and second parent nucleic acids to form truncated first and second parent nucleic acids whose length decreases incrementally over time; c) stopping said serial nucleotide removal at a plurality of different times to form a plurality of incrementally truncated first and second parent nucleic acids; d) linking separate incrementally truncated first parent nucleic acids to separate incrementally truncated second parent nucleic acids to form a plurality of first variant incrementally truncated hybrid nucleic acids, wherein said incrementally truncated first parent nucleic acids form the N-terminal coding sequence of each of said first variant incrementally truncated hybrid genes. - View Dependent Claims (5, 6)
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7. A method of making a plurality of second variant incrementally truncated hybrid nucleic acids comprising the steps of:
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a) providing a first and second parent nucleic acid; b) serially removing nucleotides from one or both termini of said first and second parent nucleic acids to form truncated first and second parent nucleic acids whose length decreases incrementally over time; c) stopping said serial nucleotide removal at a plurality of different times to form a plurality of incrementally truncated first and second parent nucleic acids; d) linking separate incrementally truncated first parent nucleic acids to separate incrementally truncated second parent nucleic acids to form a plurality of second variant incrementally truncated hybrid nucleic acids, wherein said incrementally truncated second parent nucleic acids form the N-terminal coding sequence of each of said second variant incrementally truncated hybrid genes. - View Dependent Claims (8, 9)
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Specification