CLONING ANDROGEN BINDING PROTEIN CDNAS IN LAMBDA

拟议研究的总体目标是建立一个 巴特勒大学分子生物学专业。这个项目是 旨在最大限度地提高大学生的参与度 学生们。该项目中使用的一些技术将是 简化形式在实验教学中的应用 实验室。拟议研究的目标是：1) 分离雄激素结合蛋白(ABP)亚单位的cDNA 从克隆在lambda gt11中的小鼠颌下腺cDNA和 由特定的抗ABP抗体检测，可在 首席调查员的实验室；2)获取尽可能多的不同 因为至少有三种不同的ABP，所以ABP的cDNA尽可能多 亚基可能在小鼠颌下腺中表达；3) 利用获得的ABPcDNAs来研究 在各种哺乳动物中发现了类似于ABP的基因。在完成的工作中 在本提案的范围之外，分子工具(即ABP 在这个项目中开发的)将被应用于研究 ABP基因结构及其表达方式的研究 性限唾液模式(SSP)--一种受男性影响的唾液模式 改变其中一个ABP亚基表达的基因。 拟议研究的与健康相关的意义包括 扩展我们对哺乳动物唾液蛋白的理解 发现他们在口语中的功能和作用的最终目标 健康。这些研究还可能提供信息，将 有助于最终理解生物作用(S) 非受体类固醇结合蛋白在哺乳动物中的表达。 在巴特勒建立一个分子生物学项目将 加强医学专业毕业生的背景， 牙科和研究生院。 这项工作中使用的许多方法，例如cDNA生产 和在lambda gtll中的克隆，可以用商业 可用的套件。这将极大地提高工作效率 在有限的资源和时间内进行研究 在像巴特勒这样的教学机构提供。一种程序 小鼠颌下腺组织Poly(A+)RNA的提取 腺体已经被开发出来，将被用来 积累生产cdna所需的RNA。 资助期开始。总部基地的cDNAs将由 非自动测序的双脱氧链终止法 已在印第安纳大学医疗中心以消费者为基础 中间。总部基地的cDNA将被用于搜索相关的 其他啮齿动物及其相关物种基因组DNA中的序列 使用标准Southern杂交和杂交的哺乳动物 程序。

The overall goal of the proposed research is to establish a molecular biology program at Butler University. This project is designed to maximize the involvement of undergraduate students. some of the techniques used in the project will be applied in simplified form to experiments in the teaching laboratory. The objectives of the proposed research are: 1) to isolate cDNAs for subunits of androgen binding protein (ABP) from mouse submaxillary gland cDNAs cloned in lambda gtll and detected by a specific anti-ABP antibody available in the Principal Investigator's laboratory; 2) to obtain as many distinct ABP cDNAs as possible because at least three different ABP subunits are probably expressed in mouse submaxillary glands; 3) to use the ABP cDNAs obtained to explore the occurrence of Abp-like genes in various mammalian species. In work done beyond the scope of this proposal, the molecular tools (i.e., ABP cDNAs) developed in this project will be applied to studies of Abp gene structure and to investigating the mode of expression of Sex-limited saliva pattern (Ssp), a testoster-one-influenced gene which alters the expression of one of the ABP subunits. The health-related significance of the proposed studies includes extending our understanding of mammalian salivary proteins with the ultimate goal of discovering their functions and roles in oral health. These studies may also provide information which will contribute to an eventual understanding of the biological role(s) of nonreceptor steroid binding proteins in mammals. Establishing a molecular biology program at Butler will strengthen the background of graduates who go on to medical, dental and graduate schools. Many of the methods used in this work, e.g. cDNA production and cloning in lambda gtll, can be conducted with commercially available kits. This will greatly enhance the efficiency of conduction the research with the limited resources and time available at a teaching institution such as Butler. A procedure for the isolation of poly(A+) RNA from mouse submaxillary glands has already been developed and will be used to accumulate the RNA necessary for cDNA production before the funding period starts. The ABP cDNAs will be sequenced by the dideoxy chain termination method unless automated sequencing has become available on a consumer basis at the IU medical center. The ABP cDNAs will be used to search for related sequences in the genomic DNAs of other rodents and related mammals using standard Southern blotting and hybridization procedures.