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Related Experiment Videos

Genomic analysis using a yeast artificial chromosome library with mouse DNA inserts.

J M Rossi1, D T Burke, J C Leung

  • 1Howard Hughes Medical Institute, Princeton University, NJ 08544.

Proceedings of the National Academy of Sciences of the United States of America
|March 15, 1992
PubMed
Summary

Researchers created a yeast artificial chromosome library for mouse genomic DNA, enabling efficient genome exploration. This resource facilitates chromosome walking and gene mapping, as demonstrated by mapping a region on chromosome 17.

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Area of Science:

  • Genomics
  • Molecular Biology
  • Yeast Artificial Chromosome Technology

Background:

  • Comprehensive genomic libraries are essential for gene mapping and understanding genome structure.
  • Mouse models are critical for studying human genetic diseases.
  • Yeast artificial chromosomes (YACs) allow for the cloning of large DNA fragments.

Purpose of the Study:

  • To construct a yeast artificial chromosome library containing mouse genomic DNA inserts.
  • To evaluate the library's utility for large-scale genome analysis and gene mapping.

Main Methods:

  • Construction of a yeast artificial chromosome library using mouse genomic DNA.
  • Characterization of the library for genome coverage and average insert size.
  • Development of a polymerase chain reaction (PCR)-based screening strategy using pooled DNA.

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  • Demonstration of chromosome walking capability by mapping a DNA contig.

    • Main Results:

    • The constructed library provides 2.5-fold coverage of the mouse genome.
    • Average insert size in the YAC library is 250 kilobases.
    • A 600-kilobase DNA contig surrounding the Hba-ps4 marker, linked to the fused (Fu) locus on chromosome 17, was successfully constructed.

    Conclusions:

    • The developed yeast artificial chromosome library is a valuable resource for mouse genomics.
    • The library and its screening method are effective for chromosome walking and gene mapping.
    • This resource aids in the detailed analysis of specific chromosomal regions, such as the mouse chromosome 17.