REARRANGEMENT OF VK GENES DURING ONTOGENY

免疫球蛋白可变基因是一个大的多基因家族， 成员在发育的不同时期重新排列。 年初 小鼠致癌，重链可变基因（VH）重排 仅限于位于连接点附近的几个基因 基因. 尽管对kappa光的重排知之甚少 链可变基因（VK），我们的初步数据表明，VK 基因重排比VH基因更异质 基因重排，并不限于基因的近端， 连接基因 这种重排模式之间的对比 表明可能使用不同的发育机制。 它 因此，获得更多关于VK基因信息是重要的 重新安排 本申请中提出的研究有三个目的。 (1)到 鉴定在个体发育早期重排的VK基因。 重新排列 来自三种来源的未成熟B细胞的基因将通过 测序并与VK亚家族的探针杂交。 (2)到 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。 cDNA文库将从在三个细胞中表达的mRNA构建。 发育阶段，并与基因特异性探针杂交。 (3)通过田间定位，绘制VK亚家族的染色体位置图- 反向凝胶电泳、粘粒克隆和缺失作图。 然后将来自B细胞的重排基因与它们的 在地图上的位置，以查看重排如何与 染色体定位

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.