Cloning, sequencing and characterization of a chitin synthase gene fragment from the fungus Phascolomyces articulosus /

Phascolomyces articulosus genomic DNA was isolated from 48 h old hyphae and was used for amplification of a chitin synthase fragment by the polymerase chain reaction method. The primers used in the amplification corresponded to two widely conserved amino acid regions found in chitin synthases of many fimgi. Amphfication resulted in four bands (820, 900, 1000 and 1500 bp, approximately) as visualized in a 1.2% agarose gel. The lowest band (820 bp) was selected as a candidate for chitin synthase because most amplified regions from other fimgi so far exhibited similar sizes (600-750 bp). The selected fragment was extracted from the gel and cloned in the Hinc n site of pUC19. The derived plasmid and insert were designated ^\5C\9'PaCHS and PaCHS respectively. The plasmid pUC19-PaC/fS was digested by several restriction enzymes and was found to contain BamHl and HincU sites. Sequencing of PaCHS revealed two intron sequences and a total open reading frame of 200 amino acids. The derived polypeptide was compared with other related sequences from the EMBL database (Heidelberg, Germany) and was matched to 36 other fiilly or partially sequenced fimgal chitin synthase genes. The closest resemblance was with two genes (74.5% and 73.1% identity) from Rhizopus oligosporus. Southern hybridization with the cloned fragment as a probe to the PCR reaction showed a strong signal at the fragment selected for cloning and weaker signals at the other two fragments. Southern hybridization with partially digested Phascolomyces articulosus genomic DNA showed a single band. The amino acid sequence was compared with sequences from other chitin synthase gene classes using the CLUSTALW program. The chitin synthase fragment from Phascolomyces articulosus was initially grouped in class n along with chitin synthase fragments from Rhizopus oligosporus and Phycomyces blakesleeanus which also belong to the same class, Zygomycetes. Bootstrap analysis using the neighbor-joining method available by CLUSTALW verified such classification. Comparison of PaCHS revealed conservation of intron positions that are characteristic of chitin synthase gene fragments of zygomycetous fungi.

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.

Marginal/minority popular music : the concept of the "third space" and the case for "hybridities" of cultures/identities /

The topic of this thesis is marginaVminority popular music and the question of identity; the term "marginaVminority" specifically refers to members of racial and cultural minorities who are socially and politically marginalized. The thesis argument is that popular music produced by members of cultural and racial minorities establishes cultural identity and resists racist discourse. Three marginaVminority popular music artists and their songs have been chosen for analysis in support of the argument: Gil Scott-Heron's "Gun," Tracy Chapman's "Fast Car" and Robbie Robertson's "Sacrifice." The thesis will draw from two fields of study; popular music and postcolonialism. Within the area of popular music, Theodor Adorno's "Standardization" theory is the focus. Within the area of postcolonialism, this thesis concentrates on two specific topics; 1) Stuart Hall's and Homi Bhabha's overlapping perspectives that identity is a process of cultural signification, and 2) Homi Bhabha's concept of the "Third Space." For Bhabha (1995a), the Third Space defines cultures in the moment of their use, at the moment of their exchange. The idea of identities arising out of cultural struggle suggests that identity is a process as opposed to a fixed center, an enclosed totality. Cultures arise from historical memory and memory has no center. Historical memory is de-centered and thus cultures are also de-centered, they are not enclosed totalities. This is what Bhabha means by "hybridity" of culture - that cultures are not unitary totalities, they are ways of knowing and speaking about a reality that is in constant flux. In this regard, the language of "Otherness" depends on suppressing or marginalizing the productive capacity of culture in the act of enunciation. The Third Space represents a strategy of enunciation that disrupts, interrupts and dislocates the dominant discursive construction of US and THEM, (a construction explained by Hall's concept of binary oppositions, detailed in Chapter 2). Bhabha uses the term "enunciation" as a linguistic metaphor for how cultural differences are articulated through discourse and thus how differences are discursively produced. Like Hall, Bhabha views culture as a process of understanding and of signification because Bhabha sees traditional cultures' struggle against colonizing cultures as transforming them. Adorno's theory of Standardization will be understood as a theoretical position of Western authority. The thesis will argue that Adorno's theory rests on the assumption that there is an "essence" to music, an essence that Adorno rationalizes as structure/form. The thesis will demonstrate that constructing music as possessing an essence is connected to ideology and power and in this regard, Adorno's Standardization theory is a discourse of White Western power. It will be argued that "essentialism" is at the root of Western "rationalization" of music, and that the definition of what constitutes music is an extension of Western racist "discourses" of the Other. The methodological framework of the thesis entails a) applying semiotics to each of the three songs examined and b) also applying Bhabha's model of the Third Space to each of the songs. In this thesis, semiotics specifically refers to Stuart Hall's retheorized semiotics, which recognizes the dual function of semiotics in the analysis of marginal racial/cultural identities, i.e., simultaneously represent embedded racial/cultural stereotypes, and the marginal raciaVcultural first person voice that disavows and thus reinscribes stereotyped identities. (Here, and throughout this thesis, "first person voice" is used not to denote the voice of the songwriter, but rather the collective voice of a marginal racial/cultural group). This dual function fits with Hall's and Bhabha's idea that cultural identity emerges out of cultural antagonism, cultural struggle. Bhabha's Third Space is also applied to each of the songs to show that cultural "struggle" between colonizers and colonized produces cultural hybridities, musically expressed as fusions of styles/sounds. The purpose of combining semiotics and postcolonialism in the three songs to be analyzed is to show that marginal popular music, produced by members of cultural and racial minorities, establishes cultural identity and resists racist discourse by overwriting identities of racial/cultural stereotypes with identities shaped by the first person voice enunciated in the Third Space, to produce identities of cultural hybridities. Semiotic codes of embedded "Black" and "Indian" stereotypes in each song's musical and lyrical text will be read and shown to be overwritten by the semiotic codes of the first person voice, which are decoded with the aid of postcolonial concepts such as "ambivalence," "hybridity" and "enunciation."

Righteous sounds and reproductive justice : the influence of Ani DiFranco's music for reproductive rights activists

In this thesis, I explore how the folk-rock music of Ani DiFranco has influenced the activist commitments, sensibilities, and activities of reproductive rights activists. My interest in the relation of popular music to social movements is informed by the work of Simon Frith (1987, 1996a, 1996b), Rob Rosenthal (2001), and Ann Savage (2003). Frith argues that popular music is an important contributor to personal identity and the ways that listeners see the world. Savage (2003) writes that fans develop a unique relationship with feminist/political music, and Rosenthal (2001) argues that popular music can be an important factor in building social movements. I use these arguments to ask what the influence of Ani DiFranco's music has been for reproductive rights activists who are her fans. I conducted in-depth interviews with ten reproductive rights activists who are fans of Ani DiFranco's music. All ten are women in their twenties and thirties living in Ontario or New York. Each has been listening to DiFranco's music for between two and fifteen years, and has considered herself a reproductive rights activist for between eighteen months and twenty years. I examine these women's narratives of their relationships with Ani DiFranco's music and their activist experience through the interconnected lenses of identity, consciousness, and practice. Listening to Ani DiFranco's music affects the fluid ways these women understand their identities as women, as feminists, and in solidarity with others. I draw on Freire's (1970) understanding of conscientization to consider the role that Ani's music has played in heightening women's awareness about reproductive rights issues. The feeling of solidarity with other (both real and perceived) activist fans gives them more confidence that they can make a difference in overcoming social injustice. They believe that Ani's music encourages productive anger, which in turn fuels their passion to take action to make change. Women use Ani's music deliberately for energy and encouragement in their continued activism, and find that it continues to resonate with their evolving identities as women, feminists, and activists. My study builds on those of Rosenthal (2001) and Savage (2003) by focusing on one artist and activists in one social movement. The characteristics of Ani DiFranco, her fan base, and the reproductive rights movement allow new understanding of the ways that female fans who are members of a female-dominated feminist movement interact with the music of a popular independent female artist.

Transition into the teaching profession: induction and mentoring issues surrounding secondary music teachers /

The purpose of this study was to investigate the issues surrounding the transition into the teaching profession by specifically focusing on teacher induction and mentoring issues while explicitly addressing matters of concern by secondary music teachers in a large suburban school board in southern Ontario. Participants included beginning teachers with fewer than 5 years of teaching, mid career teachers with between 6 and 15 years of instruction, and experienced teachers with more than 16 years of practice. The ' processes of mentoring and inducting new teachers within the board were examined, along with their relationships between proteges, mentors, and administrators. Further, internal and external programs specifically designed and implemented for newer music teachers were scrutinized and discussed. An analysis of key documents and literature on the subject was performed, and data were collected through 16 personal interviews. The findings suggest that although the necessity of mentoring and induction processes has begun to be recognized, there exists a fundamental relationship between mentoring and induction and the effect of the professional attachments to mentoring; the institutional and administrative supports that are enabled; and essential processes and practices between mentors and proteges. Together these three arms combine to support successfiil induction and mentoring initiatives that will help ease the transition into teaching.

Drumming toward communitas : a case study of facilitated recreational music making and the Arthurian method

The phenomenon of communitas has been described as a moment 'in and out of time' in which a collective of individuals may be experienced by one as equal and individuated in an environment stripped of structural attributes (Turner, 1 969). In these moments, emotional bonds form and an experience of perceived 'oneness' and synergy may be described. As a result of the perceived value of these experiences, it has been suggested by Sharpe (2005) that more clearly understanding how this phenomenon may be purposefully facilitated would be beneficial for leisure service providers. Consequently, the purpose of this research endeavor was to examine the ways in which a particular leisure service provider systematically employs specific methods and sets specific parameters with the intention of guiding participants toward experiences associated with communitas or "shared spirit" as described by the organization. A qualitative case study taking a phenomenological approach was employed in order to capture the depth and complexity of both the phenomenon and the purposefiil negotiation of experiences in guiding participants toward this phenomenon. The means through which these experiences were intentionally facilitated was recreational music making in a group drumming context. As such, an organization which employs specific methods of rhythm circle facilitation as well as trains other facilitators all over the world was chosen purposely for their recognition as the most respectable and credible in this field. The specific facilitator was chosen based on high recommendation by the organization due to her level of experience and expertise. Two rhythm circles were held, and participants were chosen randomly by the facilitator. Data was collected through observation in the first circle and participant- observation in the second, as well as through focus groups with circle participants. Interviews with the facilitator were held both initially to gain broad understanding of concepts and phenomenon as well as after each circle to reflect on each circle specifically. Data was read repeatedly to drawn out patterns which emerged and were coded and organized accordingly. It was found that this specific process or system of implementation lead to experiences associated with communitas by participants. In order to more clearly understand this process and the ways in which experiences associated with communitas manifest as a result of deliberate facilitator actions, these objective facilitator actions were plotted along a continuum relating to subjective participant experiences. These findings were then linked to the literature with regards to specific characteristics of communitas. In so doing, the intentional manifestation of these experiences may be more clearly understood for ftiture facilitators in many contexts. Beyond this, findings summarized important considerations with regards to specific technical and communication competencies which were found to be essential to fostering these experiences for participants within each group. Findings surrounding the maintenance of a fluid negotiation of certain transition points within a group rhythm event overall were also highlighted, and this fluidity was found to be essential to the experience of absorption and engagement in the activity and experience. Emergent themes of structure, control, and consciousness have been presented as they manifested and were found to affect experiences within this study. Discussions surrounding the ethics and authenticity of these particular methods and their implementation has also been generated throughout. In conclusion, there was a breadth as well as depth of knowledge found in unpacking this complex process of guiding individuals toward experiences associated with communitas. The implications of these findings contribute in broadening the current theoretical as well as practical understanding as to how certain intentional parameters may be set and methods employed which may lead to experiences of communitas, and as well contribute a greater knowledge to conceptualizing the manifestation of these experiences when broken down.

Educators' attitudes to philosophies of music education

This study used Q methodology to measure the extent to which individuals with five educational roles (student teacher, elementary music teacher, principal, high school music teacher, and music consultant) held five proposed philosophies of music education (hedonic, utilitarian, aesthetic cognitivism, aesthetic formalist, and praxial). Twenty-seven sUbjects participated in the Q study. These subjects were a convenience sample based on their educational role, accessibility, and willingness to participate. Participants completed a background sheet which indicated their background in music, and their responsibility for teaching music. The sUbjects in this Q study rank-ordered a set of 60 Q sort items (each item representing a proposed philosophical position) twice: Sort P to reflect current practice, and Sort I to reflect the ideal situation. The results of the sorting procedures were recorded by the participant on the response page which organized the rankings according to an approximated normal distribution as required by Q methodology. The analysis of the data suggested that the comparison across philosophical positions was significant and that the results of the interaction between philosophical position and educational role were significant, although educational role alone was not significant. Post-hoc analysis of the data was used to determine the significant differences between the levels of the, independent variables used in the model: philosophical position, educational role, and music background. A model of the association of the five philosophical positions was presented and discussed in relation to the Q study results. Further research could refine the Q sort items to better reflect each philosophical position.

Locating and sequencing of the E3 and neighbouring regions in bovine advenovirus type 2

Adenoviruses are nonenveloped icosahedral shaped particles. The double stranded DNA viral genome is divided into 5 major early transcription units, designated E1 A, E1 B, and E2 to E4, which are expressed in a regulated manner soon after infection. The gene products of the early region 3 (E3), shown to be nonessential for viral replication in vitro, are believed to be involved in counteracting host immunosurveillance. In order to sequence the E3 region of Bovine adenovirus type 2 (BAV2) it was necessary to determine the restriction map for the plasmid pEA48. A physical restriction endonuclease map for BamHl, Clal, Eco RI, Hindlll, Kpnl, Pstt, Sail, and Xbal was constructed. The DNA insert in pEA48 was determined to be viral in origin using Southern hybridization. A human adenovirus type 5 recombinant plasmid, containing partial DNA fragments of the two transcription units L4 and L5 that lie just outside the E3, was used to localize this region. The recombinant plasmid pEA was subcloned to facilitate sequencing. The DNA sequences between 74.8 and 90.5 map units containing the E3, the hexon associated protein (pVIII), and the fibre gene were determined. Homology comparison revealed that the genes for the hexon associated pV11I and the fibre protein are conserved. The last 70 amino acids of the BAV2 pV11I were the most conserved, showing a similarity of 87 percent with Ad2 pV1I1. A comparison between the predicted amino acid sequences of BAV2 and Ad40, Ad41 , Ad2 and AdS, revealed that they have an identical secondary structure consisting of a tail, a shaft and a knob. The shaft is composed of 22, 15 amino acid motifs, with periodic glycines and hydrophobic residues. The E3 region was found to consist of about 2.3 Kbp and to encode four proteins that were greater than 60 amino acids. However, these four open reading frames did not show significant homology to any other known adenovirus DNA or protein sequence.

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.

The interpretation of music videos : an interactive relationship between medium and viewer

There is a great deal of evidence to support the examination of an interactive relationship between the medium and the viewer in the interpretation of mainstream media. The exact nature of this relationship, however, is not well understood. The current study was carried out to assess the variables that may help explain why certain people interpret media, such as music videos, differently than others. Jensen's concept of reception analysis describes the relationship between the medium and the audience, and thus remains a strong focus within this study. Differences in the interpretation of music videos were investigated as a function of Absorption, gender role, screen size, age and viewing experience. Multiple regression analyses uncovered independent predictions of sexuality and violence scores by absorption and experience, as well as an interaction between absorption and screen size in the sexuality rating of the music videos.

Metal music as critical dystopia : humans, technology and the future in 1990s science fiction metal

Metal Music as Critical Dystopia: Humans, Technology and the Future in 1990s Science Fiction Metal seeks to demonstrate that the dystopian elements in metal music are not merely or necessarily a sonic celebration of disaster. Rather, metal music's fascination with dystopian imagery is often critical in intent, borrowing themes and imagery from other literary and cinematic traditions in an effort to express a form of social commentary. The artists and musical works examined in this thesis maintain strong ties with the science fiction genre, in particular, and tum to science fiction conventions in order to examine the long-term implications of humanity's complex relationship with advanced technology. Situating metal's engagements with science fiction in relation to a broader practice of blending science fiction and popular music and to the technophobic tradition in writing and film, this thesis analyzes the works of two science fiction metal bands, VOlvod and Fear Factory, and provides close readings of four futuristic albums from the mid to late 1990s that address humanity's relationship with advanced technology in musical and visual imagery as well as lyrics. These recorded texts, described here as cyber metal for their preoccupation with technology in subject matter and in sound, represent prime examples of the critical dystopia in metal music. While these albums identify contemporary problems as the root bf devastation yet to come, their musical narratives leave room for the possibility of hope , allowing for the chance that dystopia is not our inevitable future.

You get what you pay for : independent music and Canadian public policy

The aim of this MA thesis is to demonstrate how corporate concentration within the global music industry specifically affects the Canadian music industry's ability to compete for its own national audience as well as audiences worldwide. Federal public policies, regulatory regimes and subsidies are considered within the context of the structure of the global marketplace which is, in effect, an oligopoly controlled by four major corporations. Through an extensive literature review of political economy theory, Canadian public policies and music studies, as well as personal interviews conducted with Canadian musicians, entrepreneurs and public servants, I will situate my research within the body of political economy theory; present a detailed report of the structure of the global music industry; address the key players within the industry; describe the relationship between the major corporations and the independent companies operating in the industry; discuss how new technologies affect said relationships; consider the effectiveness of Canadian public policies in safeguarding the national music industry; and recommend steps that can be taken to remedy the shortcomings of Federal policies and regulatory regimes.

Steeltown scene : genre, performance and identity in the alternative independent music scene in Hamilton, Ontario

This thesis examines the independent alternative music scene in the city of Hamilton, Ontario, also known, with reference to its industrial heritage, as "Steeltown." Drawing on the growing literature on the relationship between place and popular music, on my own experience as a local musician, direct observation of performances and of venues and other sites of interaction, as well as ethnographic interviews with scene participants, I focus on the role of space, genre and performance within the scene, and their contribution to a sense of local identity. In particular, I argue that the live performance event is essential to the success of the local music scene, as it represents an immediate process, a connection between performers and audience, one which is temporally rooted in the present. My research suggests that the Hamilton alternative music scene has become postmodern, embracing forms of "indie" music that lie outside of mainstream taste, and particularly those which engage in the exploration and deconstruction of pre-existing genres. Eventually, however, the creative successes of an "indiescene" permeate mass culture and often become co-opted into the popular music mainstream, a process which, in turn, promotes new experimentation and innovation at the local level.

Bradley Institute of Music fonds

1.23 m. textual records, 1 col. post card, 1 b&w post card, 116 col. photographs, 59 b&w photographs, 6 negatives, 1 metal logo, 2 photo cuts, 7 woodcuts, 1 VHS tape, 1 stamp/press, 1 guest book, 5 account books

Leaf from a Liturgical Music Manuscript for Holy Saturday

The text reads as follows: nomen Domini invocabo. V. Date magnitudinem Deo nostro: Deus, vera opera eius, et omnes viae eius indicia. V. Deus fidelis, in quo….. Translated: I will invoke the name of the Lord. V. Give ye magnificence to our God: The works of God are perfect, and all his ways are judgments. V. Let my doctrine gather as the rain…..