Publication Date

2011

Document Type

Honors Thesis

Department

Biochemistry

Keywords

Filariasis-Prevention, Filarial worms-Genetics, Nematodes-Genetics, Nematodes as carriers of disease, Thioredoxin, Peroxidase, Molecular genetics, Gene expression, Gene regulation, Tpx-2, Brugia Malayi, Promoter

Abstract

Filarial parasitic infections continue to impact millions of lives and communities around the world, especially in tropical and subtropical regions. Most of the countries in these regions also represent some of the poorest nations in the world, and these diseases continue to have major socio-economic impacts on affected communities. The cost of treatment, low adherence to treatment strategies due to side effects and recent emergence of drug resistance are factors that have challenged current treatment strategies. Therefore, there is urgent need for novel strategies to combat the disease. Recently, several projects have been embarked upon to provide an avenue for comprehensive study of the organisms that cause these infections, which have largely remained under-studied. In order to develop better therapeutic regimes and vaccines, it is important to gain better understanding of elements that are important in the expression and regulation of various genes that help this organism to survive and escape immunological responses in two different hosts; the mosquito and human being. The antioxidant, thioredoxin peroxidase is one of the most heavily studied genes of interest as it functions as part of the mechanism for parasitic defense against radical oxygen species that are released by host organisms. In this project, an attempt has been made to characterize the BmTpx2 gene promoter by studying the effects of the deletion of the splice leader acceptor site and the 10 nucleotides upstream of this region on the activity of this promoter. The region upstream of the BmTpx2 was successfully amplified out using primers designed from the upstream region of the BmTpx2 homolog in O. volvulus, and the first exon of the BmTpx2. The PCR product has been successfully cloned into the pGL3- basic vector, albeit, in the wrong direction. Therefore, the standard protocols for ligation and transformation, which Adeola Awodele 10 are important steps in the production of these clones, have been modified in order to obtain maximal efficiency and get the desired colonies, and in the right orientation. The BmTpx2 gene warrants extensive work as an understanding of this gene, which is upregulated at the infective stage of the parasite, will contribute to increased understanding of the infective capabilities and survival strategies of filarial parasites" (30).

Language

English

Comments

69 p. : ill. (some col.) Honors project-Smith College, Northampton, Mass., 2011. Includes bibliographical references (p. 66-69)

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