Construction and characterization of bacterial artificial chromosome library of Sorghum bicolor

Sung sick Woo, Jiming Jiang, Bikram S. Gill, Andrew H. Paterson, Rod A Wing

Research output: Contribution to journalArticle

333 Citations (Scopus)

Abstract

The construction of representative large insert DNA libraries is critical for the analysis of complex genomes. The predominant vector system for such work is the yeast artificial chromosome (YAC) system. Despite the success of YACs, many problems have been described including: chimerism, tedious steps in library construction and low yields of YAC insert DNA. Recently a new E.coli based system has been developed, the bacterial artificial chromosome (BAC) system, which offers many potential advantages over YACs. We tested the BAC system in plants by constructing an ordered 13 440 clone sorghum BAC library. The library has a combined average insert size, from single and double size selections, of 157 kb. Sorghum inserts of up to 315 kb were isolated and shown to be stable when grown for over 100 generations in liquid media. No chimeric clones were detected as determined by fluorescence in situ hybridization of ten BAC clones to metaphase and interphase S.bicolor nuclei. The library was screened with six sorghum probes and three maize probes and all but one sorghum probe hybridized to at least one BAC clone in the library. To facilitate chromosome walking with the BAC system, methods were developed to isolate the proximal ends of restriction fragments inserted into the BAC vector and used to isolate both the left and right ends of six randomly selected BAC clones. These results demonstrate that the S.bicolor BAC library will be useful for several physical mapping and map-based cloning applications not only in sorghum but other related cereal genomes, such as maize. Furthermore, we conclude that the BAC system is suitable for most large genome applications, is more 'user friendly' than the YAC system, and will likely lead to rapid progress in cloning biologically significant genes from plants.

Original languageEnglish (US)
Pages (from-to)4922-4931
Number of pages10
JournalNucleic Acids Research
Volume22
Issue number23
DOIs
StatePublished - Nov 25 1994
Externally publishedYes

Fingerprint

Bacterial Artificial Chromosomes
Sorghum
Chromosomes
Chromosome
Yeast Artificial Chromosomes
Clone Cells
Libraries
Clone
Genes
Genome
Yeast
Zea mays
Organism Cloning
Probe
Cloning
Maize
Chromosome Walking
Plant Genes
Chimerism
DNA

ASJC Scopus subject areas

  • Statistics, Probability and Uncertainty
  • Applied Mathematics
  • Health, Toxicology and Mutagenesis
  • Toxicology
  • Genetics(clinical)
  • Genetics

Cite this

Construction and characterization of bacterial artificial chromosome library of Sorghum bicolor. / Woo, Sung sick; Jiang, Jiming; Gill, Bikram S.; Paterson, Andrew H.; Wing, Rod A.

In: Nucleic Acids Research, Vol. 22, No. 23, 25.11.1994, p. 4922-4931.

Research output: Contribution to journalArticle

Woo, Sung sick ; Jiang, Jiming ; Gill, Bikram S. ; Paterson, Andrew H. ; Wing, Rod A. / Construction and characterization of bacterial artificial chromosome library of Sorghum bicolor. In: Nucleic Acids Research. 1994 ; Vol. 22, No. 23. pp. 4922-4931.
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