CYTOCHROME P-450 POLYMORPHISM

我们将对兔的结构、功能和表达进行表征 人微体细胞色素P-450S 1、3b及相关形式的P-450s K和L 三群兔子。这些菌落代表近交系III/J， B/J，并选择在表型上不同的异种兔 浓度升高的P-450的表达1.抗P-450的单抗 这些酶将被用作特定的抑制剂，以评估 这个高度多样化的P-450酶家族的每个成员在 微粒新陈代谢。将采用免疫定量技术来 估计这些酶在未经处理的人群中的表达差异 动物和后代之间的交配是由基因决定的 兔子的品系。表达载体中cDNA的分子克隆将 与免疫学特征相结合以提供对 相关的cDNA克隆。这些DNA将被依次用来表征 编码这个P-450酶家族的mRNAs的数量和结构。 在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 针对该病毒的新的单抗的研制 这些P-450的特性。抗体和cdna探针将是 用来描述P-450调节的可遗传差异 在三个群体的兔子和杂交动物中的表达。

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.