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.

Wind Power Suitability Analysis for the Gulf of Maine

This map shows one option for a viable energy source that is clean, free and endless: wind power. This map shows that the coast of Maine has the potential space and wind speed to be a location for wind farms. Four NOAA buoys placed in different locations along the Maine coast are the source of the wind speed data for this project. The average wind speed of every ten minutes of every day for the year 2004 were averaged so that each buoy was represented by one number of wind speed measured in meters/ second. The values in between these four buoys were estimated, or interpolated, using ArcGIS. Other factors that I took into consideration during this lab were distance from airports (no wind farm can be with in a three mile radius of an airport ) and distance from counties (no one wants an offshore wind farm that obstructs their view). I calculated the most appropriate locations for a wind farm in ArcGIS, by adding these three layers. The final output shows an area along Mt. Desert to be the most appropriate for development.