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.

ESTIMATED NUMBER AND LOCATION OF FUTURE MOOSE-VEHICLE COLLISIONS (MVC) IN MAINE

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Estimated Number and Location of Future Moose-Vehicle Collisions (MVC) in Maine

Moose (Alces alces) are a keystone herbivore in Maine. Because of the large number of rural roads in Maine, there is a high rate of moose-vehicle collisions (MVCs), which is increasing. On-road encounters with animals resulted in 231 fatalities in the United States in 1999. Because of the fatality of MVCs, it is important to know where they are most likely to occur. I used GIS analysis to estimate where future MVCs would occur, factoring in the variables of land cover suitability for moose, distance from water bodies, locations of past MVCs, and speed limits on the roads. I ran four different analyses, each one weighting the variables equally. I also ran a regression to determine if increasing road speed was associated with the increase in the number of MVCs per length of road. There was not a strong positive relationship between the number of MVCs per length of road and the speed limit, but it was interesting to note that there were more MVCs per length of road on 35mph and 40mph roads than on 45, 50, 55 or 65mph roads. Future research on MVCs would benefit from the inclusion of include moose population density and road traffic data.

Analysis of Future Construction Possibilities on the Colby College Campus

With the recent construction of Colby Green and the current plans for the construction of several new buildings, the total area for future development on campus has declined. The goal of this study was to illustrate existing campus development and to determine where future growth could occur. GIS was used to in determining the different soil systems on campus, the current use of the land, and the boundaries of the Colby property. The project shows what potential obstacles the college will have in attempting to expand the campus and proposes where the best options are for construction are.