Abstract

Malaria is a protozoan disease caused in humans by four different species of the genus Plasmodium (P. falciparum, P. vivax, P. ovale, P. malarie) and in rodents by Plasmodium yoelii yoelii. It has been reported that 1.5 to 3 million deaths occur worldwide due to malaria and the DALY (Daily affected life years) reports about 0.76% of world population affected by the disease in some of the major countries like Africa, Asia, Latin America etc., Due to the development of resistance to drugs by the parasite, there is an urgent need and prime importance for the development of an effective vaccine against malaria. During its entire life cycle, the plasmodium sp. expresses various stagespecific proteins that are considered potential candidates for vaccine development; the major ones belong to the (i) sporozoite, (ii) erythrocytic, (iii) gametocytic stages. Merozoite surface protein 1 (MSP1) is expressed on the surface of the parasite during the erythrocytic stage, which is considered as a potential vaccine candidate. The C-terminal portion of MSP1 is considered to be an effective vaccine candidate from inhibiting the parasite invasion into RBC. PyMSP119 has been expressed in plants via the chloroplast transformation. The site specific integration of PyMSP119 gene within chloroplast genome was confirmed by PCR using specific primers and the percentage of homoplasmy vs. heteroplasmy was confirmed by Southern blot. The expression of chloroplast-derived PyMSP119 plants was confirmed by western blot using anti- PyMSP119 antibodies. These experiments showed a 17kDa protein under reducing conditions. The expression levels of PyMSP119 protein varied within transgenic plants were up to ~2% of total soluble protein (TSP) within mature leaves. To test the functionality of chloroplast-derived PyMSP119 protein, mice were immunized with the enriched chloroplast-derived PyMSP119 protein with Freund's adjuvant. The immune response of anti- PyMSP119 antibodies were tested against standard PyMSP119 protein and it yielded 1:7000 IgG titers. The immunized mice were challenged with P.yoelii infected red blood cells (35-40% parasitemia) and the percentage parasitemia suggested an inverse correlation with the immune titers. However, the concrete conclusions can be made when the study is extended to a larger animal group.

Graduation Date

2006

Semester

Fall

Advisor

Daniell, Henry

Degree

Master of Science (M.S.)

College

Burnett College of Biomedical Sciences

Department

Molecular Biology and Microbiology

Format

application/pdf

Identifier

CFE0001340

URL

http://purl.fcla.edu/fcla/etd/CFE0001340

Language

English

Release Date

October 2018

Length of Campus-only Access

None

Access Status

Masters Thesis (Open Access)

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