PIDDLE : PCR-based, in-frame, directional DNA ligation and expression

PIDDLE is a cloning strategy coupling antibody techniques to PCR. Degenerate, multifunctional header-primers derived from peptide data prime a single- or double-step PCR on genomic or cDNA template. Terminal restriction sites are thereby created to allow efficient, in-frame, directional cloning into...

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Bibliographic Details
Published in:Biotechnology techniques 1993-02, Vol.7 (2), p.91-98
Main Authors: HUGHES, J. E, QORONFLEH, M. W, BISZTRAY, G, PRATT, L. H
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Escherichia coli
Fundamental and applied biological sciences. Psychology
Genetic engineering
Genetic technics
Methods. Procedures. Technologies
Vectors (cloning, transfer, expression). Insertion sequences and transposons
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Summary:PIDDLE is a cloning strategy coupling antibody techniques to PCR. Degenerate, multifunctional header-primers derived from peptide data prime a single- or double-step PCR on genomic or cDNA template. Terminal restriction sites are thereby created to allow efficient, in-frame, directional cloning into the pBluescript-like pIDDLE phagemid. A chimeric protein is then expressed in E. coli and recognised by antibodies to the native protein. PIDDLE may also be used to generate antibodies. We describe PIDDLE cloning of phy from oat.
ISSN:0951-208X
1573-6784
DOI:10.1007/BF00157377