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Construction of a set of Saccharomyces cerevisiae vectors designed for recombinational cloning by Vincent Van Mullem; Maxime Wery; Xavier De Bolle; Jean Vandenhaute is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.

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## Abstract The Gateway^TM^ technology is becoming an increasingly popular method for cloning ORFs by recombination. It allows the transfer of any ORF flanked by specific recombination sites into any vectors harbouring the corresponding sites. Here we describe the construction of a set of 20 __Saccharomyces cerevisiae__ Gateway**TM** compatible vectors. These plasmids bear an __URA3__ or __TRP1__ selection marker. They are designed for expression without tag sequence or for C‐ or N‐terminal protein tagging with 3HA (haemagglutinin), 13MYC, 4TAP (tandem affinity purification) or GST (glutathione S‐transferase) epitopes. The centromeric vectors allow expression of DNA sequence in yeast under tetracycline‐regulatable promoters, while expression from the high copy vectors is driven by __PGK__ promoter. To test their applicability, the genes encoding the RNA polymerase I subunit Rpa12p or the TFIIS transcription factor were cloned in these vectors. Their expression was demonstrated using Western blotting or complementation assays. Copyright © 2003 John Wiley & Sons, Ltd.

Who reads Construction of a set of Saccharomyces cerevisiae vectors designed for recombinational cloning?

Author

Vincent Van Mullem; Maxime Wery; Xavier De Bolle; Jean Vandenhaute

Publisher

John Wiley and Sons; Wiley (Blackwell Publishing); John Wiley & Sons Inc.; [Chichester]; [New York]: John Wiley & Sons, ©1985-; Wiley; Hindawi Limited (ISSN 0749-503X)

Published

2003

Language

EN

Field

Biochemistry, Genetics and Molecular Biology (Life Sciences)