Abstract
Oligonucleotide synthesis was coupled with amplification by the polymerase chain reaction to generate an exact translational fusion between a plant signal sequence and an animal structural gene. A synthetic 111-mer oligonucleotide representing less than two percent of the reaction products was successfully amplified by using short primers containing restriction sites designed for ease of cloning and providing in-frame fusion. The method overcomes the length-versus-yield dilemma in oligonucleotide synthesis, and is generally adaptable to the construction of a translationally competent coding sequence from any two DNA fragments.
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Abbreviations
- ANF:
-
atrial natriuretic factor
- PCR:
-
polymerase chain reaction
- PR-5:
-
class 5 pathogenesis-related protein, also known as “PR-S”
- ssDNA:
-
single-stranded DNA
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Liew, O.W., Bullock, D.W. An alternative method for the synthesis of tailor-made genes. Plant Mol Biol Rep 14, 23–32 (1996). https://doi.org/10.1007/BF02671899
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DOI: https://doi.org/10.1007/BF02671899