Overexpression and Purification of the Oat (Avena Fatua) Protein AFN1 and Construction of a pMAL/AFN1 Expression Plasmid

Abscisic acid (ABA) is an important phytohormone with regulatory roles in many physiological processes. ABA expression is induced by environmental stresses such as drought and it is known to be an inhibitor of seed germination. A wild oat (Avena fatua) called AFN1 has been hypothesized to initiate the early stages of germination as its mRNA accumulates in nondormant seed embryos during imbibition. The polypeptide sequence of AFN1 suggests that it is an ABA glucosyl transferase. Glucosylation by AFN1 and thereby inactivation of ABA could lead to seed germination. In order to understand the role of AFN1 in germination, an ample quantity of AFN1 polypeptide is needed to test for enzymatic ABA glucosylase activity. My work has been to overexpress recombinant AFN1containing a (His)6 tag using a pRSETC E.coli expression system followed by Purification of the AFN1 protein by means of a nickel-affinity column that bind to the (His)6 tag. Due to the insufficient yield of AFN1 fusion protein obtained with this procedure, another method using a pMAL-c2x vector is now being employed. The pMAL expression system provides a method for expressing and purifying protein by tagging proteins with maltose-binding protein (MBP). It is anticipated that MBP tag will be advantageous as it can make the fusion protein more soluble and thereby yield a larger quantity of protein. Currently, work is underway on the construction of pMAL/AFN1 plasmid.

The Cultural Construction of the Maine Sporting Camps

Maine sporting camps were a cultural and social phenomenon of the urban upper and middle class. They originated in Maine in the late 1870s and early 1880s and reached their zenith around the turn of the century with over 160 in operation in eight of the sixteen counties in Maine in 1906. The period from 1880 until World War I can be considered the 'golden era' of the Maine sporting camps. After the war, with technological advancements such as the outboard motor, the proliferation of the automobile, and the introduction of a road system into rural Maine, the camps underwent significant change that warrants an entirely different cultural analysis. A number of elements came together to produce a cultural atmosphere permissive of sporting camp creation in Maine. These include changing national views upon nature, health, sport and the leisure time in which to pursue them. In the late nineteenth century, with the rise of large industrially based cities, overcrowding fostered crime and disease. An upper and middle class emerged that desired escape and separation from the lower classes. Maine was chosen for such an escape because it offered, through sporting camps, a chance to "get back to nature," by pursuing the healthful activities of hunting and fishing. At the same time these urban sportsmen and sportswomen distinguished themselves on the social Hierarchical scale from the rural inhabitants. What happened in rural Maine during the period between 1880 and World War I was the introduction of a new cultural order on the landscape. Coming primarily from urban centers on the East Coast of the United States were men, women and children who looked to Maine for vacations. These vacations were designed to put them in touch with nature by pursuing healthful activities, especially those of fishing and hunting. Coming from an environment that emphasized social standing, they ensured that these trips would perpetuate this hierarchy. They experienced nature through the Maine sporting camps, which provided them with the services and skills necessary to experience it while enjoying a degree of luxury that they were accustomed to in the urban world. The Maine sporting camps were a cultural manifestation of the urban upper and middle class, the groups that the camps were established to serve. Despite this the camps did not represent a structural duplication of urban society. Instead, the camps represented a cultural construction that was produced by interaction between members of two different conceptual and physical worlds, the blending of which, on a social level, was determined by urban mentality and rural knowledge. In the production of a cultural world meaningful to the clients, the rural world of the Maine woods was altered to meet their needs. It was not a one-sided process, however, as the clients were forced to acknowledge the importance of the rural inhabitants on the basis of their value to the clients.