CLONING ANDROGEN BINDING PROTEIN CDNAS IN LAMBDA

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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.