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The mutagenized strains were developed via transposon insertional mutagenesis. The mini- transposons (mTn) are initially introduced into a plasmid library of yeast genomic DNA, followed by transformation of the disruption alleles into a heterozygous diploid strain of yeast. Once introduced into the yeast, the transposons integrate into the corresponding genomic locus by homologous recombination. The insertion of the transposon into the open reading frame (ORF) of the gene typically disrupts gene function, or insertion upstream or downstream of the ORF may result in mis-expression of the gene.
The 6 kb mTn contains the reporter gene lacZ, flanked by lox sites and a 3xHA tag. The lacZ gene is lacking both its promoter and start codon so the expression is dependent on the transposon being inserted in-frame into a gene and subsequently transcribed and translated. Using the site-specific DNA recombinase, Cre, the 6 kb insert can be reduced to a 93-codon read-through element containing the 3X HA-tag (see HA-Tagged Yeast Collection).
The mTn technology has been used fohttp:r large-scale functional analysis of the yeast genome. Access to the resulting gene data, protocols and publications are available at the Yale Genome Analysis Center website at http://ygac.med.yale.edu/triples/triples.htm. You can obtain information on the insertion site, ORFs affected, and strains available by querying on our website by ORF ID. Background strain = Y800 (diploid).
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. We are 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.
In most cases, the insertion disrupts gene function. Some insertions, however, may not result in gene disruptions. Additionally, individual researchers should confirm that any phenotype observed is actually linked to the indicated insertion.
- P. Ross-Macdonald et al., Large-scale analysis of the yeast genome by transposon tagging and gene disruption. Nature. 402(6760), 413-418 (25 November 1999).
- A. Kumar et al., High-throughput methods for the large-scale analysis of gene function by transposon tagging. Methods Enzymol. 328, 550-574 (2000).
- A. Kumar et al., TRIPLES: a database of gene function in Saccharomyces cerevisiae. Nucleic Acids Res. 28(1), 81-84 (1 January 2000).