dc.contributor.author | Apola, Billy T | |
dc.date.accessioned | 2013-05-20T16:06:23Z | |
dc.date.available | 2013-05-20T16:06:23Z | |
dc.date.issued | 1998 | |
dc.identifier.citation | M.Sc (Biochemistry) | en |
dc.identifier.uri | http://erepository.uonbi.ac.ke:8080/xmlui/handle/123456789/23987 | |
dc.description | Master of Science Thesis | en |
dc.description.abstract | The search for glucose transporter in T.brucei was based upon the
fact that the bloodstream forms (BF)of the parasite are highly
dependent onglucose uptake for their energy metabolism. Previous
work has shown that the BF of T.brucei take up glucose across the
plasma membrane by a facilitated diffusion mechanism. The
hypothesis put forth in this project, was that the putative
gene(s) could be cloned using the Polymerase Chain Reaction
(PCR), by taking advantage of the high degree of sequence
homology within the conserved stretches of amino acid regions of
glucose transporters from prokaryotes and eukaryotes, including
a closely related trypanosomatid Leishmania. Oligonucleotide
primers constructed from the evolutionary conserved regions of
such genes, were used in a PCR using either the genomic DNA or
cDNA templates of BF and procyclic (PC) forms. The PCR products
as well as the oligonucleot4des from the conserved regions were
used as probes to screen T. brucei BF and PC lambda gtlO cDNA
librries. Several clones were isolated following the screening
exercise. The PCRproduotsand the positive clones were subcloned
into M13 phage and partially-sequenced using Sanger's dideoxy
chain termination method. None had glucose transporter orglucose
transporter related sequences after computer homology searches.
However some cDNA clones isolated after screening with the PCR .
products generated from genomic DNA templates, had open reading
frames having homology to co.lLaqeri of other organisms. The
homology is highest in Herpesvirus.collagen-like protein which
has 62.4% nucleotide identity. The results of the study showed
that the primers constructed were unable to isolate glucose
transporter(s) In the trypanosoma brucei, but may be suitable to
isolate genomic sequences having collagen like sequences. | en |
dc.description.sponsorship | University of Nairobi | en |
dc.language.iso | en | en |
dc.title | A search for glucose transporters in trypanosoma brucei leads to identification of collagen like DNA sequences | en |
dc.type | Thesis | en |
local.publisher | Department of Chemistry, University of Nairobi | en |