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Purification and Characterization of Trypanolusin form the Midgut of Desert Locust (Schistocerca gregaria), A Non-Vector of Trypanosomes

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dc.contributor.author Mwaura, John Gitau
dc.date.issued 2008-09
dc.date.accessioned 2022-09-26T13:35:03Z
dc.date.available 2022-09-26T13:35:03Z
dc.description.abstract African trypanosomosis. caused by protozoan parasites of the genus T/jtpunosoma. is a serious threat to human and animal health and thus a major hindrance to economic development in sub-Saharan Africa. TI:\t]7CI}’1l)SOI77(l undergoes complex life cycle that is marked by differentiation and multiplication in the tsetse (Glossina Spp.) vector and in the vertebrate host. Under field and experimental conditions, most flies are refractory to infection by trypanosomes. The establishment of infection in the tsetse midgut is an important first step in the disease transmission. However. the tsetse inidgut presents a hostile environment that kills most of the trypanosomes ingested during an infective blood meal. Some of the tsetse midgut factors involved in this interaction include lectins_ trypsins. trypsin-like molecules and trypanolysins among which trypanolysins have received least attention. Trypanolysins' presence and functions in haematophagous and non-haematophagous non-vector insects is not well documented. The present study was initiated to study trypanolysin properties in non-vector insect locust (Sc‘hi¢'I0cercu gregariu). Trypanolysin was isolated from the midgut of locust S. gregariu through several consecutive chromatographic steps namely; gel filtration on Sephadex G-150. DEAE anion exchange and Concanavalin A Sepharose affinity chromatography. Further purification was achieved by electro-elution from native Polyacrylamide gel electrophoresis (PAGE). Trypanolysin eluted at the major peak on the gel filtration column, bound and eluted at about 260mM salt concentration on DEAE anion-exchange column but eluted in the unbound fraction in Con A affinity column. The native molecular weight was estimated to be about 28 KDa while under SDS-PAGE a single band of approximately 29 l\'Da was obtained. The proteineous nature of the trypanolysin was evidenced by the denaturation by temperatures above 50°C. Trypanolysin activity was not significantly affected by divalent cations (Mg 2+ and Cay) suggesting non-metallo co-factor dependence. The locust midgut trypanolysin is not glycosylated as evidenced by its failure to stain in periodic acid Schiff stain. The midgut trypanolysin elicited weak immunogenic response in a na'i've New Zealand White rabbit. Western blot analysis using polyclonal antisera against the trypanolysin show/ed no cross reactivity with midgut protein extracts from haematophagous (tsetse. mosquitoes and sand fly) and non-haematophagous (cockroach. housefly) insects indicating that its S’. greguriu specific. The findings of this work indicate that S. gregariu midgut trypanolysin exhibits significant differences in physical and chemical characteristics from trypanolysins in the vector (ilossina xpp. en_US
dc.language.iso en en_US
dc.publisher Egerton University en_US
dc.subject Trypanolusin -- Midgut -- Desert Locust -- Non-Vector en_US
dc.title Purification and Characterization of Trypanolusin form the Midgut of Desert Locust (Schistocerca gregaria), A Non-Vector of Trypanosomes en_US
dc.type Thesis en_US

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