Construction and characterization of a yeast artificial chromosome library of Arabidopsis which is suitable for chromosome walking View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1991-05

AUTHORS

Erwin Grill, Chris Somerville

ABSTRACT

A yeast artificial chromosome (YAC) genomic library of Arabidopsis thaliana was constructed in a derivative of the vector pYAC4 which was modified to facilitate the production of end-specific probes for chromosome walking. Experiments in which a subset of 2300 clones from the library were probed with 30 restriction fragment length polymorphism (RFLP) markers indicated that, on the average, the entire genome is represented once in each 800 YAC clones. Thus, the complete library of more than 20000 YACs is expected to contain most or all of the Arabidopsis genome with a high probability. The YAC clones examined in the sample had an average insert size of approximately 150 kb±10 and represented more than 5% of the Arabidopsis genome. Based on the properties of the library and the currently available RFLP maps for Arabidopsis, only one or two steps from flanking RFLPs should be sufficient to isolate an average gene in Arabidopsis by chromosome walking with the YACs. In order to facilitate chromosome walking, a method for the production of hybridization probes from the ends of the inserts was employed that is based on a combination of pre-amplification of the vector/insert junction sequences via the polymerase chain reaction and specific transcription from T3 or T7 RNA polymerase promoters flanking the cloning site in the YAC vector. More... »

PAGES

484-490

Identifiers

URI

http://scigraph.springernature.com/pub.10.1007/bf00260662

DOI

http://dx.doi.org/10.1007/bf00260662

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1046378691

PUBMED

https://www.ncbi.nlm.nih.gov/pubmed/1674816


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