CYTOCHROME P-450 POLYMORPHISM

我们将对兔抗肿瘤基因的结构、功能和表达进行研究， 微粒体细胞色素P-450 s 1，3b和P-450 s K和L的相关形式， 三只兔子。 这些菌落代表近交系III/J， B/J，和选择的远系繁殖兔，不同的表型在 表达升高浓度的P-450 1. 单克隆抗体 这些酶将被用作特异性抑制剂， 这个高度多样化的P-450酶家族的每个成员在 微粒体代谢 免疫定量技术将用于 估计这些酶的表达在未处理的 动物之间的遗传定义之间的交配后代 兔子的品种。 在表达载体中cDNA的分子克隆将 结合免疫学表征，以提供 相关cDNA克隆。 这些cDNA将依次用于表征 编码这个P-450酶家族的mRNA的数量和结构。 在用所述融合蛋白转化的微生物中表达的融合蛋白， 含有编码这些酶的cDNA的表达载体将用于 开发额外的单克隆抗体， 这些P-450的特征。 抗体和cDNA探针将被 用于表征P-450调节中的遗传差异 在兔和远交动物的三个群体中的表达。

We will characterize the structure, function and expression of rabbit microsomal cytochroms P-450s 1, 3b and related forms of P -450s K and L in three colonies of rabbits. These colonies represent inbred strains III/J, B/J, and selected outbred rabbits that differ phenotypically in the expression of elevated concentrations of P-450 1. Monoclonal antibodies to these enzymes will be used as specific inhibitors in order to assess the role of each member of this highly diverse family of P-450 enzymes in microsomal metabolism. Immunoquantitative techniques will be employed to estimate differences in the expression of these enzymes among untreated animals and among progeny of matings between the genetically defined strains of rabbits. Molecular cloning of cDNAs in expression vectors will be coupled with immunological characterization to provide identification of pertinent cDNA clones. These cDNAs will be used in turn to characterize the number and structure of mRNAs encoding this family of P-450 enzymes. the fusion proteins expressed in microorganisms transformed with the expression vectors containing cDNAs encoding these enzymes will be used in the development of additional monoclonal antibodies for the characterization of these P-450s. The antibodies and cDNA probes will be used to characterize heritable differences in the regulation of P-450 expression among the three colonies of rabbits and outbred animals.