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One-step PCR mediated strategy for the construction of conditionally expressed and epitope tagged yeast proteins.

ABSTRACT: With the availability of the complete yeast genomic sequence, techniques which allow the rapid functional analysis of genes of interest are of increasing importance. Here we report a technique which allows the initial characterisation of genes of interest, through the construction of conditionally expressed mutations for functional analyses and the generation of epitope-tagged fusion proteins for immuno-localisation and immuno-purification, entirely by PCR.

SUBMITTER: Lafontaine D

PROVIDER: S-EPMC146098 | biostudies-other | 1996 Sep

REPOSITORIES: biostudies-other

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One-step PCR mediated strategy for the construction of conditionally expressed and epitope tagged yeast proteins.

Lafontaine D D Tollervey D D

Nucleic acids research 19960901 17

With the availability of the complete yeast genomic sequence, techniques which allow the rapid functional analysis of genes of interest are of increasing importance. Here we report a technique which allows the initial characterisation of genes of interest, through the construction of conditionally expressed mutations for functional analyses and the generation of epitope-tagged fusion proteins for immuno-localisation and immuno-purification, entirely by PCR. ...[more]

PMID: 8811105

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