Rapid one-step recombinational cloning.

Nucleic Acids Research
Changlin FuBrian Hauge

Abstract

As an increasing number of genes and open reading frames of unknown function are discovered, expression of the encoded proteins is critical toward establishing function. Accordingly, there is an increased need for highly efficient, high-fidelity methods for directional cloning. Among the available methods, site-specific recombination-based cloning techniques, which eliminate the use of restriction endonucleases and ligase, have been widely used for high-throughput (HTP) procedures. We have developed a recombination cloning method, which uses truncated recombination sites to clone PCR products directly into destination/expression vectors, thereby bypassing the requirement for first producing an entry clone. Cloning efficiencies in excess of 80% are obtained providing a highly efficient method for directional HTP cloning.

References

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Nov 15, 2000·Genome Research·J L HartleyM A Brasch
Jun 20, 2002·Protein Expression and Purification·Lynda DieckmanFrank R Collart

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Citations

May 1, 2009·Plant Physiology·Songbiao ChenGuo-Liang Wang
Feb 1, 2012·Microbial Cell Factories·Antoine GruetChristophe Bignon
Feb 7, 2014·Molecular Biotechnology·Quirino ScheferCarsten Grötzinger
Oct 3, 2014·Nucleic Acids Research·Andrew B KennedyChristina D Smolke
Apr 1, 2015·The Journal of Cell Biology·Kyle M DraheimDavid A Calderwood
Dec 20, 2015·The FEBS Journal·Antoine GruetChristophe Bignon
Jan 11, 2016·American Journal of Human Genetics·Tychele N TurnerEvan E Eichler
Jan 29, 2011·Chembiochem : a European Journal of Chemical Biology·Misha V GolynskiyMaarten Merkx
Jan 27, 2011·The Plant Journal : for Cell and Molecular Biology·Till IschebeckIngo Heilmann
Dec 13, 2016·The Plant Journal : for Cell and Molecular Biology·Anna Ophelia MüllerTill Ischebeck
Mar 12, 2021·Plant Science : an International Journal of Experimental Plant Biology·Nadav EisnerDudy Bar-Zvi

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Methods Mentioned

BETA
two
new hybrid
PCR
electrophoresis

Software Mentioned

DNAstar

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