The cloning and reconstitution of a bovine adenovirus type 2 E3 deletion mutant and the sequencing and analysis of the early 4 region

Recombinant Adenoviruses (Ads) have been shown to have potential applications in three areas: gene therapy, high level protein expression and recombinant vaccines.' At least three different locations within the Ad genome can be deleted and subsequently used for the insertion of foreign sequences. These include the Early 3 (E3), Early 1 (E1) and Early 4 (E4) regions. Viral vectors of this type have been well studied in Human Ads 2 and 5, however one has not yet been constructed for Bovine Adenovirus Type 2 (BAV2). The E3 region is located between 76.6 and 86 m.u. on the r-strand and is transcribed in a rightward direction. The gene products of the Early 3 region (E3) have been shown to be non-essential for viral replication, in vitro, but are required for host immunosurveillance. This study represents the cloning and reconstitution of a BAV2 E3 deletion mutant. A deletion of 1800bp was made within the E3 region of BAV2 and the thymidine kinase gene was subsequently inserted in the deleted area . . The plasmid pdlE3-4tk1 (23.4Kbp) was constructed and used to to facilitate homologous recombination with the wild type BAV2 to produce a mutant. Southern Blotting and Hybridization results suggest the presence of a BAV2 E3 deletion mutant with thymidine kinase sequences present. The E4 region of Human Adenovirus types 2 and 5 is located at the extreme right end of the genome (91.3 map units - 99.1 map units) and is transcribed in a leftward direction giving rise to a complicated set of differentially spliced mRNAs. Essentially there are 7 open reading frames (ORFs) encoding for at least 7 polypeptides. The gene products encoded by the E4 region have been shown to be essential for the expression of late viral genes, host cell shutoff and normal viral growth. We have cloned and sequenced the right end segment between 90.5 map units and 100 map units of the BAV2 genome. The results show several open reading frames which encode polypeptides exhibiting homology to three polypeptides encoded by the E4 region of human adenovirus type 2. These include the 14kDa protein encoded by ORF1, the 34kDa protein encoded by ORF6 and the 13kDa protein encoded by ORF3. The nucleotide sequence, restriction enzyme map, and ORF map of the E4 region could be very useful in future molecular manipulation of this region and could possibly explain the slow growth rate of BAV2 in MDBK cells.

Toward a grounded theory of trigger events and leadership development in early adulthood

The purpose of this research study was to develop a conceptual model through the use of a grounded theory approach, which explains how trigger events are related to leadership development. Trigger events are experience that cause developmental growth and may result in an increased ability to lead (Luthans and Avolio (2003). In this study, there were two phases of data collection. First participants completed the Washington University Sentence Completion Test (WUSCT), where their respective leadership developmental stage was measured. Second, participants were involved in two in-depth interviews where an understanding was reached as to how various trigger events have impacted their leadership development. From these data, a conceptual model was developed to explain the relationship between trigger events and leadership development. Participants described trigger events as being important developmental periods, during which time they grew as people and became more capable leaders.

Early Education Foundation: Portuguese Canadian Mothers' Preferences and Choices

The purpose of this study was to explore Portuguese-Canadian mothers' preferences and choices regarding their children's early care and education. The findings revealed that Portuguese-Canadian mothers value early care and education and are conscious of their role in their children's lives. Regardless of the type of care setting, the participants' responses revealed that the caregiver's care, emotion, and responsiveness are most important. More than developing "savvy" children, we need to nourish "happy" children. The study's participants include 9 Portuguese Canadian mothers without any assumption of a hyphenated identity and who have moved away from their immigrant parents' script. They embraced the vision of their children's success and cultivated their vast potential. Their responses revealed that the family, culture, and traditions are important factors in their child's academic and social growth and played a critical role in establishing the foundations for learning. The research study findings showed that the field of early care and education is undergoing a paradigm shift and that other practices, ideologies, and theories are surfacing. This study aimed to help develop a new grounded theory that contributes to a better understanding of this arena. The present findings reveal important issues for further discussion and lay a theoretical and empirical framework for future research in early education and care.