NON-TUMORIGENIC TERATOCARCINOMA

越来越清楚的是，某些基因，取决于 他们表达的时间和性质，可以调节有些对立 肿瘤发生和分化的过程。胚胎学 癌症(EC)细胞系统在研究中已被证明具有特殊的价值 这两个进程之间的关系以及它们之间的相互关联性。 因此，在试图识别新的调控基因时，申请人 通过逆转录病毒插入诱变EC细胞系并产生 与EC细胞在致瘤性和致瘤性方面都不同的突变株 差异化。申请人已经证明，突变是由 插入到半合子位置的单个插入。对两个序列进行排序 该地点紧邻的5‘和3’侧翼区域已显示开放 阅读框与任何已知的小鼠基因没有同源性。这表明 突变的表型可能是由于病毒插入而出现的 转化为一种新的鼠标编码序列。申请人建议进一步使用 侧翼序列信息、动物园杂交、CpG岛分析和外显子 捕捉技术以识别位于该点或其附近的转录单位 轨迹。该转录本将用于分离全长cdna克隆。 然后对cdna和基因组克隆的基因进行比较和测序。 并检查5‘监管区域。申请者还将分析 转录本在细胞中表达的功能后果 转染体和转基因小鼠并测定组织和胚胎 特定的表达模式。最后，她会孤立和刻画 其表达因以下原因而改变的特定蛋白质 基因破坏。申请人相信这些研究将会 结果发现了一种新的小鼠基因，并对我们的 对控制两者的分子和细胞因素的理解 增殖分化。

It has become increasingly clear that certain genes, depending upon the time and nature of their expression, can regulate the somewhat opposed processes of both tumorigenesis and differentiation. The embryonal carcinoma (EC) cell system has proven to be of particular value in studies of these two processes as well as in evaluating their interrelatedness. Therefore, in an attempt to identify novel regulatory genes the applicant mutagenized an EC cell line via retroviral insertion and generated a mutant line that differs from EC cells in both tumorigenicity and differentiation. The applicant has shown that the mutation was created by a single insertion into a hemizygous site. Sequencing of both the immediate 5' and 3' flanking regions at this site has revealed open reading frames with no homology to any known mouse gene. This indicates that the mutant phenotypes may have arisen as a result of viral insertion into a novel mouse coding sequence. The applicant proposes to use further flanking sequence information, zoo blotting, CpG island analysis and exon trapping techniques to identify a transcription unit at or near this locus. This transcript will be used to isolate a full length cDNA clone and then compare and sequence the gene from both cDNA and genomic clones and examine the 5' regulatory region. The applicant will also analyze the functional consequences of expression of the transcript in cell transfectants and transgenic mice and determine the tissue and embryonic specific expression patterns. Finally, she will isolate and characterize specific proteins whose expression has been altered as a consequence of the gene disruption. The applicant believes that these studies will result in the identification of a novel mouse gene and contribute to our understanding of the molecular and cellular elements controlling both proliferation and differentiation.