TERATOCARCINOMA AND EMBRYONAL TUMORS: SURFACE ANTIGENS

T/t复合体在遗传控制中具有重要作用， 正常早期发育中的细胞定型。 几 重要的新研究使隐性致死突变 分子分析：（1）其中两个已经 在充分表征的MHC内映射; 12在H-2D之间 和TL，并且w5在遗传上与H-2K不可分离。 (2)185 kb 的H-2K区域的-单倍型已被克隆， 限制性酶切图谱。 初步结果显示， 包含几个早期胚胎表达序列。 (3)的 相关细胞表面抗原gp 87的结构基因具有 两个物理映射的副本，一个在Qa区域，一个在 两个H-2K基因之间的联系 我们打算利用这一最近的 试图识别和结构化描述的信息 有趣的早期胚胎表达基因，包括w5和12。 我们提出的总体方法是：1）搜索H-2K 利用cDNA序列对胚胎表达基因进行区域克隆 胚胎癌细胞，并使用独特的拷贝探针， 基因组互补序列，以探测来自 正常的早期胚胎 为了明确识别w5， 我们建议将其H-2K区域结构与 回复 2)第二种方法是从结构上研究 单倍型的D-TL区域的组织，其中12个基因 所在 我们将使用CHEF凝胶分析和现有的 重组体以细化12的图谱位置。 3)最后我们将 继续进行相关糖蛋白的分子分析 抗原“糖蛋白87”的检索和测序克隆 cDNA文库。 我们将学习有趣的表达方式 使用多种技术鉴定基因，包括原位 杂交到胚胎的切片， 突变体转基因胚胎拯救 我们的长远目标是 了解分子机制和遗传控制， 正常早期发育中的细胞定型， 胚胎性肿瘤的非定型。

The T/t-complex has an important role in the genetic control of cellular commitment in normal early development. Several important new studies make the recessive -lethal mutations more accessible to molecular analysis: (1) two of them have been mapped within the well characterized MHC; 12 is between H-2D and TL, and w5 is genetically inseparable from H-2K. (2) 185 kb of the H-2K region of -haplotypes has been cloned in cosmids and restriction mapped. Preliminary results indicate this region contains several early embryo-expressed sequences. (3) the structural gene for a -associated cell surface antigen, gp87, has two copies physically mapped, one in the Qa region and one between the two H-2K genes. We intend to use this recent information to try to identify and structurally characterize interesting early embryo-expressed genes including w5 and 12. The overall approaches we propose are: 1) to search the H-2K region cosmids for embryo expressed genes using cDNA from embryonal carcinoma cells, and to use unique copy probes from the genomic cosmids to probe available cDNA libraries from normal early embryos. In an effort to specifically identify w5, we propose to compare its H-2K region structure to that of a revertent. 2) The second approach is to structurally study the organization of the D-TL region of -haplotypes where the 12 gene is located. We will use CHEF gel analysis and existing recombinants to refine the map position of 12. 3) Finally, we will continue the molecular analysis of the -associated glycoprotein antigen "glycoproteins 87" by retrieving and sequencing clones from cDNA libraries. We will study the expression of interesting genes identified using a variety of techniques including in situ hybridization to cut sections of embryos and in the case of the mutants, transgenic embryo rescue. Our long term objective is to understand the molecular mechanisms and genetic control of cellular commitment in normal early development, and noncommitment in embryonal tumors.