VH REGION DIVERSIFICATION AMONG HAPLOTYPES

描述：（改编自研究者摘要）长期目的是 为了了解VH区在细胞中多样化的潜在机制， 人免疫球蛋白重链基因复合物及其作用 人类健康的多样化。工作假设是， 这一地区多样化的结果是显著的生物学 和遗传的影响，并可能包含“脚印”留下的遗传 事件负责多样化。对这些脚印的研究可能会有所帮助 了解负责VH区域多样化的机制。的 VH区是高度多样化的。据信，没有单倍型含有 同一组VH基因片段。因此，即使一个完整的核苷酸序列 对于通过使用二倍体材料产生的VH区，序列可能不 代表任何单倍型中的实际序列，并且远远不足以 了解VH区的物理结构。在研究报告中，50 具有高度多样化遗传背景的男性人类受试者将被 招募的供体单倍型中VH基因片段的数量和组成 将通过分析来自这些供体的单个单倍体精子细胞来确定。 将根据结果选出四名有代表性的捐助者。单倍型 将确定这四个受试者中VH基因片段的组织 通过分析单个精子的单个DNA片段由此产生的信息 将用于组织来自噬菌体的含有VH基因片段的克隆 这些捐赠者的基因组文库。所有VH基因区段的序列和 它们的侧翼区含有启动子，转录调节元件， 并确定每个单倍型中的重组信号序列。的 将检查每个单倍型的详细物理结构，以确定 限制性酶切作图后的遗传目的区域。的序列 将确定确定的区域。由此产生的数据将用于 推断其余46个单倍型中的VH基因组织 个人和进一步分析VH区域多样化的影响 以及多样化背后的机制。年制定的办法 该研究也可用于分析其它基因单倍型基因组 多基因家族，包括那些参与免疫系统发育的家族， 系统为了确定染色体的详细物理结构 具有单拷贝序列的区域。

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.