REARRANGEMENT OF VK GENES DURING ONTOGENY

个体发育过程中 VK 基因的重排

• 批准号: 3301930
• 负责人: PATRICIA J GEARHART
• 金额: $ 17.13万
• 依托单位: JOHNS HOPKINS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-01-01 至 1991-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3301930
• 关键词: B lymphocyte; antibody formation; gel electrophoresis; gene expression; gene rearrangement; genetic library; genetic manipulation; genetic mapping; growth /development; immunoglobulin genes; laboratory mouse; messenger RNA; molecular cloning; nucleic acid hybridization; nucleic acid sequence; tissue /cell culture

Immunoglobulin variable genes are a large, multigene family whose members rearrange at different times in development. Early in mouse oncogeny, rearrangement of heavy chain variable genes (VH) is limited to a few genes that are located proximal to the joining genes. Although less is known about rearrangement of kappa light chain variable genes (VK), our preliminary data indicates that VK gene rearrangements are more heterogeneous than VH gene rearrangements and are not restricted to genes proximal to the joining genes. This contrast between the rearrangement patterns suggests that different developmental mechanisms may be used. It is therefore important to obtain more information on VK gene rearrangements. The research proposed in this application has three aims. (1) To identify VK genes that rearrange early in ontogeny. Rearranged genes from three sources of immature B cells will be examined by sequencing and hybridization to probes of VK subfamilies. (2) To determine if VK and VH gene expression correlates with the programmed appearance of antibody specificities during ontogeny. cDNA libraries will be constructed from mRNA expressed at three stages of development and hybridized with gene-specific probes. (3) To map the chromosomal position of VK subfamilies by field- inversion gel electrophoresis, cosmid cloning and deletion mapping. Rearranged genes from B cells will then be compared to their position on the map to see how rearrangement correlates with chromosomal location.

免疫球蛋白可变基因是一个大的多基因家族，其 成员在发展的不同时期进行重新安排。早些时候 小鼠致癌、重链可变区基因(VH)重排 仅限于位于连接近端的几个基因 基因。尽管人们对卡帕光的重新排列知之甚少 链可变基因(VK)，我们的初步数据表明VK 基因重排比VH基因更具异质性 重排，并不限于基因的近端 加入基因。这种重排模式之间的对比 这表明可以使用不同的发育机制。它 因此，获取更多关于VK基因的信息是重要的 重新安排。 本申请中提出的研究有三个目的。(1)至 识别个体发育早期重排的VK基因。重新排列 来自三个来源的未成熟B细胞的基因将通过 对VK亚家族的探针进行测序和杂交。(2)至 确定VK和VH基因表达是否与 个体发育过程中抗体特异性的程序化出现。 将从在三个基因上表达的mRNA构建cDNA库 并与基因特异性探针杂交。 (3)定位VK亚家族的染色体位置。 倒置凝胶电泳、粘粒克隆和缺失定位。 然后将来自B细胞的重排基因与它们的 地图上的位置，查看重新排列如何与 染色体定位。