IDENTIFYING AFR2--GENE REGULATOR IN LIVER PATHOLOGY

描述：甲胎蛋白（AFP）基因是一种癌胎儿抗原， 在发育中的胎儿和在病理条件下， 再生和肿瘤发生。 AFP基因水平 在小鼠肝再生过程中， 非连锁常染色体基因座Afr2。 近交系C57B1/6小鼠表达8至10倍 在肝再生过程中AFP低于野生型C3H/He小鼠。 AFP下限 再生肝中的表达可能反映了其他调节过程，如 C57B1/6小鼠比C3H/He小鼠对肝癌发生的易感性低 小鼠 从这些和其他遗传学研究来看，Afr2基因似乎 是多态的。 由于Afr2调节AFP表达，因此使用这些不同的 小鼠品系提供了一个独特的机会，以确定一个基因调节 肝再生过程中AFP的表达。 使用来自鼠标的信息 基因组计划，在第一系列的实验中，Afr2基因将被 基因图谱 该基因座的克隆DNA将从 酵母人工染色体文库。 转录序列将被分离 通过外显子捕获和/或cDNA的定向选择。 两个标准将是 用于确定转录序列以及 先前定位到该位点的基因是Afr2：与Afr2基因连锁 肝再生过程中的基因和表达。 述第二触发器串列中 在实验中，将制备全长Afr2 cDNA克隆，转染 进入原代肝细胞培养系统，并测试其 调节AFP基因表达。 分析两种已知的 将比较Afr2等位基因。 鉴定和分离 Afr2基因将为研究植物与人之间的关系提供一个工具。 发育生长、受控生长和不受控的致瘤生长 在肝脏中。

DESCRIPTION: The alpha-fetoprotein (AFP) gene is an onco-fetal antigen, expressed in the developing fetus and under the pathological conditions of regeneration and tumorigenesis in the adult. The level of AFP gene expression during liver regeneration in mice is regulated by a genetically unlinked autosomal locus, Afr2. Inbred C57B1/6 mice express 8 to 10 fold less AFP during liver regeneration than wild type C3H/He mice. Lower AFP expression in regenerating liver may reflect other regulatory processes, as C57B1/6 mice are less susceptible to liver carcinogenesis than are C3H/He mice. From these and other genetic studies, it appears that the Afr2 gene is polymorphic. Since Afr2 regulates AFP expression, use of these different mouse strains provides a unique opportunity to identify a gene regulating AFP expression during liver regeneration. Using information from the Mouse Genome Project, in the first series of experiments, the Afr2 gene will be genetically mapped. Cloned DNA of this locus will be identified from a Yeast Artificial Chromosome library. Transcribed sequences will be isolated by exon-trapping and/or direction selection of cDNAs. Two criteria will be used to determine the likelihood that the transcribed sequences, as well as genes previously mapped to the locus, are Afr2: genetic linkage to the Afr2 gene and expression during liver regeneration. In the second series of experiments, a full length Afr2 cDNA clone will be prepared, transfected into a primary hepatocyte culture system, and tested for its ability to regulate AFP gene expression. Analysis of the activity of the two known Afr2 alleles will be compared. Identifying and isolating a clone of the Afr2 gene will provide a tool to study the relationships between developmental growth, controlled growth, and uncontrolled tumorigenic growth in the liver.