Project Details


DESCRIPTION: (Adapted from investigator's abstract) The long-term objective is
to understand the mechanisms underlying diversification of the VH region in the
human immunoglobulin heavy chain gene complex and the effect of the
diversification on human health. The working hypothesis is that the differences
resulted from the diversification in this region are of significant biological
and genetic implications, and may contain "footprints" left by the genetic
events responsible for the diversification. Study of these footprints may help
to understand the mechanisms responsible for the VH region diversification. The
VH region is highly diversified. It is believed that no haplotypes contain the
same set of VH gene segments. Therefore, even if a complete nucleotide sequence
for the VH region is generated by using diploid materials, the sequence may not
represent the actual sequences in any haplotypes and is far from sufficient for
understanding physical structure of the VH region. In the proposed study, 50
male human subjects with highly diversified genetic backgrounds will be
recruited. VH gene segment number and composition in the donors' haplotypes
will be determined by analyzing single haploid sperm cells from these donors.
Four representative donors will be selected based on the results. The haplotype
organization of the VH gene segments in these four subjects will be determined
by analyzing single DNA fragments from single sperm. The resulting information
will be used to organize the VH gene segment-containing clones from phage
genomic libraries for these donors. The sequences of all VH gene segments and
their flanking regions containing promoters, transcription regulatory elements,
and recombination signal sequences in each haplotype will be determined. The
detailed physical structure of each haplotype will be examined to identify the
regions of genetic interest after restriction mapping. The sequences of the
identified regions will be determined. The resulting data will be used to
deduce the VH gene organization in the haplotypes of the remaining 46
individuals and for further analysis impact of the VH region diversification
and the mechanisms underlying the diversification. The approaches developed in
the study may also be used to analyze haplotype gene organization of other
multi-gene families including those involved in the development of the immune
system. And to determine the detailed physical structure of the chromosomal
regions with single copy sequences.
Effective start/end date8/1/996/30/04


  • Genetics
  • Molecular Biology


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