random, in-frame affinity tags created with minitransposons
- Generate HA epitope tagged proteins for
immunolocalization or immunoprecipitation using antibodies against HA.
- A potential source of conditional alleles and
hypomorphic mutants (generated by transposon insertion of 3X HA Tag).
- Find strains and insertion sites of the HA epitope tag via our user friendly on-line query.
The HA-Tagged Yeast Collection contains over 2,400 yeast mutagenized
strains each producing a single protein with an inserted triple
hemagglutinin epitope tag (3xHA)*. HA tags are useful for a variety of
functional studies including immunolocalization, immunoprecipitation
and analysis of binding sites using immunodetection. Insertion via
mini- transposon (mTn) mutagenesis allows for tagging within the coding
region of the protein in a non-biased fashion thus resulting in the
tagging of both annotated and previously unidentified ORFs1.
The HA insertion potentially provides several means by which yeast
protein function may be studied. HA-tagged yeast proteins have been
used effectively in large-scale studies of protein localization;
full-length HA tagged proteins can be localized to the appropriate
cellular region where they can be detected using an antibody to the HA
epitope2. Native amounts of protein can easily be purified from
individual yeast strains using immunoprecipitation with a commercially
available HA antibody. Also, HA insertion may potentially generate
conditional alleles and hypomorphic mutants that exhibit partial gene
function of particular importance in the analysis of essential genes.
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| We provide certain clone resources developed by leading academic laboratories. Many of these resources address the needs of specialized research communities not served by other commercial entities. In order to provide these as a public resource, we depend on the contributing academic laboratories for quality control. Therefore, these are distributed in the format provided by the contributing institution "as is" with no additional product validation or guarantee. Thermo Fisher Scientific is not responsible for any errors or performance issues. Additional information can be found in the product manual as well as in associated published articles (if available). Alternatively, the source academic institution can be contacted directly for troubleshooting. |
Construction:
The HA-Tagged yeast collection utilizes mini-transposon (mTn)
constructs and shuttle mutagenesis to develop a collection of
mutagenized yeast genes for analysis of expression patterns,
localization, and phenotypic analysis1,2. The constructs originally
contained a 6-kb multipurpose transposon containing β- galactosidase
reporter sequence, selection markers for both E. coli and
Saccharomyces, flanked by lox sites and the required transposon
elements. Following reduction by Cre-lox recombination, a 93-codon
region consisting primarily of the read through 3xHA tag remains. The
mTn technology has been used for large-scale functional analysis of the
yeast genome. Public access to the resulting gene data, protocols, and
publications is available at the Yale Genome Analysis Center3.
Background strain = Y800 (diploid).
*Strains are not guaranteed to produce a fully functional protein and complete analysis is to be performed by the individual researcher. Information on strains that have been successfully immunolocalized is available on the TRIPLES website.
The HA-Tagged Yeast Collection is
provided in a diploid ura3 leu2 background. Individual strains are
provided as a live culture in a 2ml tube. Each tube contains YPAD broth
supplemented with 15% glycerol. Store the strain at 4°C for up to one
week or -80°C indefinitely.
Bulk orders are shipped as glycerol stocks in 96-well microtiter plates
and are a culture of the HA Tagged Yeast in YPAD media and glycerol.
These plates ship on dry ice and can be maintained indefinitely at
-80°C.
Contact our customer service department at info@openbiosystems.com or
call at 888-412-2225 for a quote and estimated shipping time on bulk
orders.
1P. Ross-Macdonald, et.al., Nature 402, 413 (1999).
2A. Kumar, et.al., Genes & Development 16, 707 (2002).
3Kumar, A., Cheung, K.-H., Ross-Macdonald, P., Coelho, P.S.R., Miller, P., and Snyder, M. Nucleic Acids Res. 28, 81 (2000).
