Normal and diseased human tissues were analyzed for the expression of genes of the carcinoembryonic (CEA) family. The use of portions of the CEA and NCA cDNA clones as hybridization probes has shown the existence of CEA and NCA transcripts in colon mucosa, NCA transcripts in breast tumours, and NCA and a third CEA-like transcript (CGM6) in chronic myelocytic leukemia (CML) cells. / A library prepared from CML cells was used to isolate a cDNA clone of CGM6. The predicted product of this novel CEA family member displays 85% sequence similarity to CEA and NCA and its expression is limited to CML and normal human bone marrow cells. / Southern blots of DNA did not reveal any CEA gene rearrangements or amplifications accompanying CEA expression. DNA digested with methylation-sensitive endonucleases demonstrated that CEA expression is correlated with a decreased level of methylation in the 5$ sp prime$ region of the CEA gene.

The long term goal of this research is to overexpress a mutant form of COII and create a dominant negative mutation in COX, transgenic mice containing the dominant negative mutation could be used to investigate the effects of the respiratory chain deficiencies which are characteristic of mitochondrial myopathies on mitochondrial biogenesis and gene expression. This thesis reports an attempt to express rat COII from a nuclear transgene. Using PCR strategies, the mitochondrial matrix targeting sequence from rat ornithine carbamyltransferase, was attached to a version of the rat COII gene which had been recoded into the universal genetic code. The transgene was cloned into the pLXSN retroviral vector and GP+E-86 ecotropic packaging cells were transfected with the construct. Infectious retrovirus particles were collected and used to infect C2C12 mouse myoblast and NIH3T3 mouse fibroblast cell lines. Expression of mRNA from the transgene was assessed in both cell lines using an RT-PCR strategy. For each cell type, protein expression from the transgene was assessed in the two clones showing the highest levels of mRNA expression. COII protein levels were determined by Western blot analysis using peptide-specific polyclonal antibodies directed to the C-terminus of rat COII. The transgene was expressed in a rabbit reticulocyte lysate system in which transcription and translation are coupled. In vitro translation products were immunoprecipitated using the COII peptide-specific antibodies. (Abstract shortened by UMI.)

The Glucocorticoid receptor (GR) is one of many steroid hormone receptors. It controls broad physiological gene networks, confers pathological effects in a range of disease states, and offers an excellent target for therapeutic intervention. Therefore, it is necessary to better understand the mechanisms of GR regulation. In particular, we are interested in better understanding the protein-nucleotide interactions (transcription factors interacting with transcription factor binding sites). Upon glucocorticoids-hormone binding, the GR forms a protein-nucleotide interaction with a specific transcription factor binding site known as a glucocorticoid response element (GRE). This research has employed three different but complementary bioinformatics approaches to identify Mouse genes putatively transcriptionally regulated by GR. Firstly, we focus on the problem of searching for putative GREs in the complete Mouse genome using a position weight matrix. This produced a large number of putative GREs. Most of these are likely false positive predictions. Secondly, two different strategies are used to improve the accuracy of our framework: combinatorial analysis of multiple TFs/modules of TFBSs and phylogenetic footprinting (PF). The number of putative GREs can be reduced by 97.9% using the module of TFBSs analysis, 97.7% using the PF analysis, and 99.9% using both module and PF analyses. In each step, a statistical test has been used to measure the significance of the results.

Methylenetetrahydrofolate reductase (MTHFR) catalyzes the reduction of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, a substrate for remethylation of homocysteine to methionine. Methionine is then converted to S-adenosylmethionine (SAM), the universal methyl donor. In the mouse, MTHFR levels are highest in the testis and mice deficient for a targeted mutation in the Mthfr gene are infertile. Analysis of prenatal gonocyte development provided evidence that the defect in the Mthfr mouse testis occurs shortly after birth. The expression profiles at the mRNA and protein levels suggest that Mthfr is expressed in the male germ line and is developmentally regulated, with a possible role in meiosis. Testicular phenotype was further characterized by altering the expression of this enzyme in an Mthfr folate deficiency animal model. While postnatal folate deficiency appears not to have adverse effects on spermatogenesis, it results in significantly reduced sperm numbers in males from the developmental study, regardless of their Mthfr genotype.

Thesis (Ph.D.)--University of Illinois at Urbana-Champaign, 2007. / Source: Dissertation Abstracts International, Volume: 68-06, Section: A, page: 2528. Adviser: Steven R. Leigh. Includes bibliographical references (leaves 136-166) Available on microfilm from Pro Quest Information and Learning.

Thesis (Ph.D.)--Indiana University, Dept. of Biology, 2007. / Source: Dissertation Abstracts International, Volume: 68-05, Section: B, page: 2790. Adviser: Susan Strome. "Title from dissertation home page (viewed Jan. 24, 2008)."

Thesis (Ph.D.)--University of California, San Francisco, 2006. / Source: Dissertation Abstracts International, Volume: 68-01, Section: B, page: 0085. Adviser: Jonathan S. Weissman.

Thesis (Ph.D.)--University of California, San Francisco, 2005. / Source: Dissertation Abstracts International, Volume: 66-12, Section: B, page: 6424. Adviser: Christine Guthrie.

Thesis (Ph.D.)--University of California, San Francisco, 2006. / Source: Dissertation Abstracts International, Volume: 68-01, Section: B, page: 0085. Adviser: Jonathan S. Weissman.

Thesis (Ph.D.)--University of California, San Francisco, 2005. / Source: Dissertation Abstracts International, Volume: 66-12, Section: B, page: 6369. Adviser: Nigel Killeen.