Integrative selection of human chromosome-specific yeast artificial chromosomes

William J. Pavan, Roger H Reeves

Research output: Contribution to journalArticle

Abstract

Human specific "integrative selection vectors" (ISVs) were designed to optimize integration of a yeast-selectable marker specifically into yeast artificial chromosomes (YACs) derived from human but not mouse DNA. ISVs were transformed into a YAC genomic library constructed from DNA of a human-mouse somatic cell hybrid containing chromosome 21 (HSA21) as the only human chromosome. One percent of the yeast in the original library contained HSA21-derived YACs; between 45% and 54% of the yeast recovered after transformation with ISV vectors contained human YACs. Integrative selection provides a rapid means of obtaining a highly enriched population of human chromosome-specific YACs by eliminating the labor-intensive steps of isolating and screening primary transformants. The procedure is biased toward the selection of YACs that contain a large number of targets for homologous recombination; thus, libraries constructed by this procedure will be composed primarily of the largest YACs in the population.

Original languageEnglish (US)
Pages (from-to)7788-7791
Number of pages4
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number17
StatePublished - 1991

Fingerprint

Yeast Artificial Chromosomes
Human Chromosomes
Yeasts
Libraries
Human Artificial Chromosomes
Chromosomes, Human, Pair 21
Genomic Library
Hybrid Cells
Homologous Recombination
DNA
Population

Keywords

  • Genetic vector
  • Homologous recombination
  • Yeast artificial chromosome library

ASJC Scopus subject areas

  • General
  • Genetics

Cite this

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AB - Human specific "integrative selection vectors" (ISVs) were designed to optimize integration of a yeast-selectable marker specifically into yeast artificial chromosomes (YACs) derived from human but not mouse DNA. ISVs were transformed into a YAC genomic library constructed from DNA of a human-mouse somatic cell hybrid containing chromosome 21 (HSA21) as the only human chromosome. One percent of the yeast in the original library contained HSA21-derived YACs; between 45% and 54% of the yeast recovered after transformation with ISV vectors contained human YACs. Integrative selection provides a rapid means of obtaining a highly enriched population of human chromosome-specific YACs by eliminating the labor-intensive steps of isolating and screening primary transformants. The procedure is biased toward the selection of YACs that contain a large number of targets for homologous recombination; thus, libraries constructed by this procedure will be composed primarily of the largest YACs in the population.

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