East Carolina University : Research and Creative Achievement Week 2011
All males were mated with normal C57Black 7-week old female mice. The offspring will be monitored for changes in developmental pattern and body weight curve. Upon reaching 5 weeks, half of the offspring will be subjected to 60% fat diet for 12 weeks and half will be used as controls on regular mouse chow. Tissue collected from fathers and offspring will be screened for genome-wide changes in DNA methylation pattern. We anticipate differences in susceptibility to diabetes between offspring of high-fat fathers and offspring of control and exercise groups. Yeast two hybrid interaction studies, Wayne Rummings, Department of Biology, East Carolina University, Greenville, NC 27858 The lab that I perform research in investigates the proteins involved in eukaryotic DNA replication using interaction studies in yeast, as well as both genetic and phenotypic analysis in Drosophila melanogaster. My research thus far has been centered around the roles of Mcm10 in replication. The protein encoded by this gene is one of the highly conserved mini-chromosome maintenance proteins (MCM) that are involved in the initiation of eukaryotic genome replication and has been shown to be involved with heterochromatin formation (Apger et al 2010). To this point, my role has been to clone eleven genes in preparation for yeast two-hybrid interaction studies with Mcm10, as well as preparing growth media for the lab, and maintaining fly stocks. This process of cloning has consisted of successfully amplifying DNA via polymerase chain reactions (PCR), verifying results through agarose gel electrophoresis, and subsequently cloning the PCR products into bacterial plasmid. These clones were then used to generate yeast two-hybrid vectors. My endeavors have provided the starting materials for many projects in the lab and have opened doors for future work for myself and other students conducting molecular genetic analysis in Drosophila. I propose to use yeast two-hybrid interaction studies to characterize the domains of Mcm10 that are responsible for interactions with the genes I have cloned. Through this process I plan to elucidate the regions responsible for these interactions at a 200aa resolution. The progression of this work will further the knowledge of the interaction networks of Mcm10, contribute to the understanding of eukaryotic replication initiation, continue to define the role of Mcm10 in replication and chromatin dynamics, as well as aiding in the basic understanding of cell proliferation and mechanisms of cancer.
P114
Regulation of Connexin 43 by Cyclic AMP in Vascular Smooth Muscle, Danielle Martin, Jonathan C. Fox, Chintamani N. Joshi, David A. Tulis, Department of Biology, East Carolina University, Greenville, NC 27858 P115 Connexin 43 (Cx43) is the predominant gap junction protein in vascular smooth muscle cells (VSMCs) and plays important roles in maintaining normal vascular function as well as in the response to disease or injury. Connexins are trans-membrane proteins that facilitate intercellular communication by regulating movement of ions and other signaling molecules, including cyclic nucleotides, through gap junction intercellular communications (GJICs). The physiologic efficacy of 192