Construction of bovine adenovirus type 3 E1 and E3, substitution plasmids and the sequencing analysis of DNA pol and pTP /

The relative ease to concentrate and purify adenoviruses, their well characterized mid-sized genome, and the ability to delete non-essential regions from their genome to accommodate foreign gene, made adenoviruses a suitable candidate for the construction of vectors. The use of adenoviral vectors in gene therapy, vaccination, and as a general vector system for expressing foreign genes have been documented for some time. In this study, the objective was to rescue a BAV3 E1 or E3 recombinant vector carrying the kanamycin resistant gene, a dominant selectable marker with useful applications in studying vectored gene expression in mammalian cells. To accomplish the objective of this study, more information about BAV3 DNA sequences was required in order to make the manipulation of the virus genome accessible. Therefore, sequencing of the BAV3 genome from 1 1 .7% to 30.8% was carried out. Analysis of the determined sequences revealed the primary structure of important viral gene products coded by E2 including BAV3 DNA pol and precursor to terminal protein. Comparative analysis of these proteins with their counterparts from human and non human adenoviruses revealed important insights as to the evolutionary lineage of BAV3. In order to insert the kanamycin resistance gene in either E1 or E3, it was necessary to delete BAV3 sequences to accommodate the foreign gene so as not to exceed the limit of the packaging capacity of the virus. To construct a recombinant BAV3 in which a foreign gene was inserted in the deleted E1 region, an E1 shuttle vector was constructed. This involved the deletion from the viral sequences a region between 1.3% to 9% and inserting the kanamycin resistance gene to replace the deletion. The E1 shuttle vector contained the left (0%- 53.9%) segment of the genome and was expected to generate BAV3 recombinants that can be grown and propagated in cells that can complement the missing E1 functions. To construct a similar shuttle vector for E3 deletion, DNA sequences extending from 78.9% to 82.5% (1281 bp) were deleted from within the E3 region that had been cloned into a plasmid vector. The deleted region corresponds to those that have been shown to be non-essential for viral replication in cell culture. The resulting plasmid was used to construct another recombinant plasmid with BAV3 DNA sequences extending from 37.1% to 100% and with a deletion of E3 sequences that were replaced by kanamycin resistance gene. This shuttle plasmid was used in cotransfections with digested viral DNA in an attempt to rescue a recombinant BAV3 carrying the kanamycin resistance gene to replace the deleted E3. In spite of repeated attempts of transfection, El or E3 recombinant BAV3 were not isolated. It seems that other approaches should be applied to make a final conclusion on BAV3 infectivity.

Implicit and explicit functions in the acquisition of a phonics rule in the word recognition of learning disabled and non learning disabled students /

One group of 12 non learning disabled students and two groups of 12 learning disabled students between the ges of 10 and 12 were measured on implicit and explicit knowledge cquisition. Students in each group implicitly cquired knowledge bout I of 2 vocabulary rules. The vocabulary rules governed the pronunciation of 2 types of pseudowords. After completing the implicit acquisition phase, all groups were administered a test of implicit knowledge. The non learning disabled group and I learning disabled group were then asked to verbalize the knowledge acquired during the initial phase. This was a test of explicit knowledge. All 3 groups were then given a postlest of implicit knowledge. This tcst was a measure of the effectiveness of the employment of the verbalization technique. Results indicate that implicit knowledge capabilities for both the learning disabled and non learning disabled groups were intact. However. there were significant differences between groups on explicit knowledge capabilities. This led to the conclusion that implicit functions show little individual differences, and that explicit functions are affected by ability difference. Furthermore, the employment of the verbalization technique significantly increased POStlest scores for learning disabled students. This suggested that the use of metacognitive techniques was a beneficial learning tool for learning disabled students.

Private versus public : how one gay professor negotiates the boundaries between his personal and professional lives /

This study examined how one university professor negotiated the boundaries between his personal life as a gay man and his professional life as a teacher. Using his sexual orientation as a focal point, the study explored the circumstances and underlying assumptions that influenced this professor's decisions to disclose information of a personal nature. Data collection was solicited from a number of sources: (a) In-depth interviews with the participant, his colleagues, students, and friends; (b) Field observation of the participant teaching over a 3 -day period; and (c) A document review of lesson plans, course outlines, student feedback forms, and the participant's teaching portfolio. The researcher maintained both observation journals and reflective journals during this process. Data analysis using the constant comparative method elicited several themes. The participant engaged in a variety of strategies in disclosing his sexual orientation that included: (a) no disclosure at all, (b) assuming people knew, (c) casually mentioning it in conversation, and (d) deliberately planning to tell someone. The participant also engaged in an ongoing assessment of his environment that included evaluating the level of risk in disclosing his sexual orientation and assessing the listener's ability to receive the information. The participant cited numerous reasons for disclosing his sexual orientation. Further inquiry revealed a number of belief systems that underlined these reasons. These belief systems included beliefs around privacy, authenticity, teaching, manners, professionalism, and homosexuality. The conclusions suggested that the participant utilized a consistent process in both his personal and professional lives to determine what information was kept private and what information was made public. While the process used to determine the degree of disclosure was consistent, the actual disclosures themselves varied widely in nature.