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Stable transformation of pleomorphic bloodstream form Trypanosoma brucei.

Research output: Contribution to Journal/MagazineJournal articlepeer-review

Published
  • P MacGregor
  • F Rojas
  • S Dean
  • KR Matthews
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<mark>Journal publication date</mark>31/08/2013
<mark>Journal</mark>Molecular and Biochemical Parasitology
Issue number2
Volume190
Number of pages3
Pages (from-to)60-62
Publication StatusPublished
Early online date5/07/13
<mark>Original language</mark>Undefined/Unknown

Abstract

African trypanosomes differentiate between various developmental stages both in mammalian hosts and their tsetse vector to adapt to and survive in the different environments they encounter. In the bloodstream, trypanosomes naturally exist as either proliferative slender-forms or non-proliferative stumpy-forms, the latter being responsible for both prolonged infection and transmission. However, most trypanosome studies are carried out on laboratory-adapted monomorphic cell lines, incapable of differentiating to stumpy-forms or completing the life cycle through the tsetse fly. Partly, this has been due to the inefficiency of transfection of pleomorphic strains which have retained the ability to generate stumpy-forms. Recently, Amaxa Nucleofector® technology was shown to increase transfection efficiency for monomorphic bloodstream forms. Using this technology we have optimised a similar method for pleomorphic bloodstream form transfection, generating transfection efficiencies of 10−7–10−6. This permits routine genetic manipulation of pleomorphic lines, which have the most biological relevance for trypanosomes in the field.