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Can I read HB tag modules for PCR‐based gene tagging and tandem affinity purification in Saccharomyces cerevisiae on EtoBox?
HB tag modules for PCR‐based gene tagging and tandem affinity purification in Saccharomyces cerevisiae by Christian Tagwerker; Hongwei Zhang; Xiaorong Wang; Liza S. Z. Larsen; Richard H. Lathrop; G. Wesley Hatfield; Bernhard Auer; Lan Huang; Peter Kaiser is a Biochemistry, Genetics and Molecular Biology article available to read on EtoBox.
What is HB tag modules for PCR‐based gene tagging and tandem affinity purification in Saccharomyces cerevisiae about?
## Abstract We have recently developed the HB tag as a useful tool for tandem‐affinity purification under native as well as fully denaturing conditions. The HB tag and its derivatives consist of a hexahistidine tag and a bacterially‐derived __in vivo__ biotinylation signal peptide, which support sequential purification by Ni^2+^‐chelate chromatography and binding to immobilized streptavidin. To facilitate tagging of budding yeast proteins with HB tags, we have created a series of plasmids with various selectable markers. These plasmids allow single‐step PCR‐based tagging and expression under control of the endogenous promoters or the inducible __GAL1__ promoter. HB tagging of several budding yeast ORFs demonstrated efficient biotinylation of the HB tag __in vivo__ by endogenous yeast biotin ligases. No adverse effects of the HB tag on protein function were observed. The HB tagging plasmids presented here are related to previously reported epitope‐tagging plasmids, allowing PCR‐based tagging with the same locus‐specific primer sets that are used for other widely used epitope‐tagging strategies. The Sequences for the described plasmids were submitted to GenBank under Accession Number
Who reads HB tag modules for PCR‐based gene tagging and tandem affinity purification in Saccharomyces cerevisiae?
It is typically read by researchers, students, and practitioners in Biochemistry, Genetics and Molecular Biology.
- Author
- Christian Tagwerker; Hongwei Zhang; Xiaorong Wang; Liza S. Z. Larsen; Richard H. Lathrop; G. Wesley Hatfield; Bernhard Auer; Lan Huang; Peter Kaiser
- 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
- 2006
- Language
- EN
- Field
- Biochemistry, Genetics and Molecular Biology (Life Sciences)