T-DNA promoter tagging in Nicotiana tabacum : a thesis presented in fulfilment of the requirements for the degree of Master of Philosophy in Genetics at Massey University, Palmerston North, New Zealand

Plant development is primarily controlled at the level of gene expression. In order to analyse this regulation it is necessary to isolate genes which are involved in organ development through cellular and tissue determination or which respond to environmental signals. Promoter tagging was chosen in order to identify genes potentially associated with plant development by their spatial and temporal pattern of expression. The introduction of a promoterless reporter gene tag allows the expression patterns of plant genes to be readily characterised. A new series of promoter tagging vectors were constructed from the plasmid pPCV604 (Koncz, 1989). The selectable kanamycin resistance marker gene from pBin6 (Bevan, 1984) was cloned into pPCV604 to create pGT. The hygromycin phosphotransferase gene in pGT was then replaced with a promoterless (β-glucuronidase (gus) gene coupled with octopine synthase termination sequence subcloned from pKiwi101a (Janssen and Gardner, 1989) creating pGTG. This binary transformation vector required the helper pRK replication functions of Agrobacterium tumefaciens strain GV3101. In order to bypass this restriction, the vector sequence of pBin19 was combined with the T-DNA of pGTG to create pBin19-GTG. The latter plasmid was found to have a higher Agrobacterium tumefaciens-mediated Nicotiana tabacum transformation efficiency in strain LBA4404 than pGTG in strain GV3101. In both the pGTG and pBin19-GTG promoter tagging vectors the promoterless gus gene has an initiation codon 62 base pairs inside the T-DNA. This sequence includes translation termination codons in all three reading frames. Therefore, insertion of the T-DNA into a plant gene could lead to activation of the gus gene, under the control of the plant gene promoter, via transcriptional fusion. Nicotiana tabacum leaf segments were transformed with pGTG or pBin19-GTG and transgenic plants selected on kanamycin. A population of 87 transgenic tobacco plants were fluorometrically screened for GUS activity in leaf and root material; 37% were found to contain GUS activity, indicating a high frequency of promoter tagging. Two transgenic plants with root specific gus expression were analysed histochemically. Progeny after self-fertilisation lacked GUS activity, though this was restored in progeny of one plant with 5-azacytidine treatment, suggesting involvement of methylation in the gene silencing. Southern hybridisation, inverse PCR cloning of T-DNA flanking sequences and segregation on kanamycin indicated the presence of multiple T-DNA copies within the primary transformants. Furthermore, inverse PCR sequence from one plant indicated multiple and truncated T-DNA insertions at one or more loci. A further population of transformed plants was generated with pBin19-GTG and histochemically screened for GUS activity in roots (14 positive from 147 tested), shoots (27 positive from 147) and floral organs (14 positive from 56). Overall, combining results from all plant organs tested, an average of 33% of plants were found with GUS activity in one or more organs. A diverse range of patterns of gus expression were observed and described including patterns involving root branching. Forty four plants from this population were analysed for T-DNA copy number via Southern hybridisation with a gus probe (right border junction T-DNA) and nptII probe (central T-DNA). Multiple copies were frequently found with an average of 3.3 T-DNA copies per transgenic plant. Overall, an average of 11% of T-DNA insertions were found to be involved in gus activation. Comparison of the fluorometric (37% positive, 87 plants tested) and histochemical (22% positive, 147 plants tested) screens for GUS activity in root and shoot material was discussed and it is suggested that further care is needed in assigning promoter tagging hits from fluorometric screening. Variable expression was observed with promoter tagged genes. It is suggested that further research is required to determine whether this variation was due to silencing, perhaps by methylation, or was a result of the tagged promoters' normal expression patterns

Effects of Manganese Chloride on Blood Formation in Rabbits

The objectives of these investigations were to determine the effect of manganese on the following: red and white blood cell counts, differential counts, packed cell volume, hemoglobin, reticulocyte composition, and manganese and iron determinations in respect to possible changes in these measurements with prolonged exposure to manganese

Life Cycle Demographic Events And Economic Well-being: A Study Of Families With Young Children In Canada, 1971-1986

This study examines change in the economic welfare of families with preschool age children in Canada over the period 1971-1986. While the purpose of this study is to document change in the economic welfare of such families through alternative indicators of economic well-being, more broadly it seeks to examine interrelationships between family change and economic well-being. This study begins with a critical review of previous literature that focuses upon the link between family and economic change, prior to moving on to a general discussion of several of the practical and conceptual difficulties that surface in the definition of economic well-being and income poverty. The public use sample tapes of the 1971, 1981 and 1986 Canadian censuses are relied upon in a descriptive overview of recent changes (across subsamples of Canadians as defined by life cycle stage) prior to a more detailed focus upon families with preschool age children.;In examining the interrelationships between family change and economic well-being for families with preschool age children, a multivariate model of the income to needs ratio is relied upon (involving both family/demographic and non-demographic variables). Through a series of regressions, the relative importance of specific factors to change in the economic well-being of families are estimated. On the basis of the proposed model, simulation procedures are also introduced in estimating the relative importance of family/demographic change to trends as observed in income poverty.;Overall, the period 1971-1986 witnessed (1) a decline in the number of children per family, (2) an upward shift in the age distribution of fertility, (3) an increased involvement of women in the paid labour force, and (4) and increased incidence of lone parenthood. Irrespective of the detrimental impact of an increased proportion of all young families headed by a female lone parent, the overall impact of family/demographic change over the period of interest appears to be one which makes a positive contribution to the economic welfare of young families. Evidence suggestive of an increased level of income inequality is also presented, as might be expected given a continuing income gap between single parent and dual income families (both of which steadily increased as a proportion of all such families over the reference period)

The role of the social foundations of education in programs of teacher preparation in Canada

This paper argues that the social foundations of education, and particularly the disciplines of history, philosophy and sociology of education, must continue to play an integral role in programs of teacher education. We report on the decline of the study of history of education within Faculties of Education in Canada as an example of the marginalization of the role of the social foundations in teacher education programs generally. In this context we furnish what we take to be some of the strongest reasons for the requirement for future teachers to engage with the social foundations—some of these arguments apply to all of the foundational areas, and some apply to specific foundational disciplines. Some of these arguments will be familiar, some new. We conclude that if a teacher education program in Canada is to be of a very high quality then it must include a strong social foundations component.Keywords: teacher education, sociology of education, history of education, philosophy of education, social foundations of educationCet article affirme que les fondements sociaux de l'éducation, et particulièrement les disciplines telles que l'histoire, la philosophie et la sociologie de l'éducation, doivent continuer à jouer un rôle intégral dans les programmes de formation des enseignants. Nous faisons rapport du le déclin de l'étude de l'histoire de l'éducation au sein de facultés d'éducation au Canada, comme un exemple de la marginalisation du rôle des fondements sociaux dans les programmes de formation des enseignants en général. Dans ce contexte, nous fournissons ce que nous pensons être parmi les plus fortes raisons, en tant qu'obligation pour les futurs enseignants, de se familiariser avec les fondements sociaux - Certains de ces arguments s'appliquent à l'ensemble des domaines fondamentaux, et certains s'appliquent seulement à des disciplines fondamentales spécifiques. Certains seront connus, d'autres nouveaux. Nous en concluons que si un programme de formation des enseignants au Canada se veut être un programme d'excellence, alors il doit inclure une composante importante sur les fondements sociaux.Mots clés: formation des enseignants, sociologie de l'éducation, histoire de l'éducation, philosophie de l'éducation, fondements sociaux de l'éducatio

Sodium Hexametaphosphate as an Aid in the Treatment of Periodontal Disease

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/66734/2/10.1177_00220345440230050301.pd

Elderly’s Perception about The Value of Assistive Technologies for their Daily Living: Impacting Factors and Theoretical Support

This paper presents a review of relevant studies found in a systematic search for research on the elderly’s perception about the value of assistive technologies. We found that the majority of papers did not use any theory to frame the research question or explain their results. We suggest that invariably occurs in benefits realization through the use of empowering technologies designed to provide training to help older people maintain their functional capabilities. Maintaining these functional capabilities is important for people to live independently for longer and in order to assess the benefits more quickly; we suggest researchers use the capability approach. The existing theories of adoption appear to be much more suited to supportive technologies that aid elderly in their functional disabilities. This is mainly due to the fact that the immediate benefits i.e. usefulness of these technologies can be seen more easily compared to empowering technologies

Suppository formulations as a potential treatment for nephropathic cystinosis.

Nephropathic cystinosis is a rare autosomal recessive disease characterised by raised lysosomal levels of cystine in the cells of all the organs. It is treated by the 6-h oral administration of the aminothiol, cysteamine, which has an offensive taste and smell. In an attempt to reduce this frequency and improve the treatment, cysteamine-containing polyethylene glycol suppositories were prepared and evaluated for dissolution and stability. The results demonstrated that cysteamine release was complete after 30 min, and that there was a uniform drug distribution within the formulations. Twelve-month stability tests highlighted a potential incompatibility among some excipients, although stability was demonstrated for the cysteamine suppositories up to 6 months. These suppositories may provide a useful alternative to the current oral therapy for cystinosis