Transformation-associated recombination (TAR) cloning of tumor-inducing Xmrk2 gene from Xiphophorus maculatus

We modified the TAR methodology of YAC clone construction for application to fish genomic DNA isolated from Xiphophorus maculatus. YAC libraries were developed using the XIR1 repeat sequence as the recombinational hook. Construction of these libraries demonstrates that Xiphophorus DNA sequences can...

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Bibliographic Details
Published in:Marine biotechnology (New York, N.Y.) N.Y.), 2001-06, Vol.3 (Supplement 1), p.S168-S176
Main Authors: Kirchner, J M, Ivanova, V, Samson, A, Noskov, V N, Volff, J N, Resnick, M A, Walter, R B
Format: Article
Language:English
Subjects:
Clones
Cloning
Construction
Deoxyribonucleic acid
DNA
Fish
Fish genetics
Gene sequencing
Genetic recombination
Genetic transformation
Genetics
Genomics
Neoplasms
Nucleotide sequence
Recombination
Regions
Replication origins
Sequencing
Xiphophorus
Xiphophorus maculatus
Xmrk gene
Yeast
Yeasts
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Summary:We modified the TAR methodology of YAC clone construction for application to fish genomic DNA isolated from Xiphophorus maculatus. YAC libraries were developed using the XIR1 repeat sequence as the recombinational hook. Construction of these libraries demonstrates that Xiphophorus DNA sequences can function as hooks in the yeast recombination system and that X. maculatus genomic DNA contains sequences that provide origin of replication function in yeast. By screening a subset of Xiphophorus YAC clones, we isolated a clone harboring the Xmrk2 locus that is associated with spontaneous and induced melanomagenesis. Modifications of the TAR technique allowed the targeted cloning of specific genes from genomic regions ranging in size from cDNAs to several hundred kilobases. Specific genomic regions can be isolated in a directional manner from fixed map locations to saturate these areas with physical markers. We discuss the applications of these and other yeast recombinational processes to fish genetics.
ISSN:1436-2228
1436-2236
DOI:10.1007/s1012601-0039-9