| Applications |
In hosts lacking a nonsense suppressor, lysogenization and lytic growth of the phage are prevented by the cI- Int- and P- phenotypes of the bacteriophage. Recovery of transpositions into plasmid replicons are obtained by transforming plasmid DNA from kanR colonies into a lambda resistant host such as E. coli NK8017 (ATCC 47057). |
| Insert |
DNA: genomic Insert lengths(kb): 9.399999618530273 Gene product: uridine monophosphate synthetase( UMP synthase, orotate phosphoribosyltransferase, orotidine 5'-phosphate decarboxylase, orotate phosphoribosyltransferase 1) [URA3] |
| Comments |
Restriction digests of the clone give the following sizes (kb): EcoRI--23.0, 19.4, 9.4, 5.6, 3.8, 3.0; BamHI--27.0, 17.0, 11.0; PvuII--22.5, 15.0, 9.4, 6.0, 3.3, 1.7, 1.1, 0.7, 0.5. In hosts lacking a nonsense suppressor, lysogenization and lytic growth of the phage are prevented by the cI- Int- and P- phenotypes of the bacteriophage. Recovery of transpositions into plasmid replicons are obtained by transforming plasmid DNA from kanR colonies into a lambda resistant host such as E. coli NK8017 (ATCC 47057). For insertional mutagenesis of bacterial or yeast genes carried in plasmid vectors. Contains the mini-Tn10-LUK insert that consists of the following features: IS10R - nt 63 - promoterless lacZ element starting at codon 8 - nt 3140 - URA3 - nt 4320 - kanR - nt 6023 - IS10L - nt 6090. |
| References |
Huisman O, et al. A Tn10-lacZ-kanR-URA3 gene fusion transposon for insertion mutagenesis and fusion analysis of yeast and bacterial genes. Genetics 116: 191-199, 1987. PubMed: 3038670
Kleckner N, et al. Uses of transposons with emphasis on Tn10. Methods Enzymol. 204: 139-180, 1991. PubMed: 1658561
Riles L, et al. Physical maps of the six smallest chromosomes of Saccharomyces cerevisiae at a resolution of 2.6-kilobase pairs. Genetics 134: 81-150, 1993. PubMed: 8514151
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