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Transfection systems for Babesia bovis: A review of methods for the transient and stable expression of exogenous genes by Carlos E. Suarez; Terry F. McElwain is a Immunology and Microbiology article available to read on EtoBox.

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With the recently sequenced Babesia bovis genome, a large pool of genes with unknown function was identified. The ability to complement and knock-out both unknown and previously identified genes would be a valuable tool to better understand gene function in B. bovis parasites. This review describes recent advances in the development of transient and stable transfection systems for B. bovis. Transient transfection constructs were initially generated using the promoter and the 3' region of the rap-1 genes of B. bovis controlling expression of luciferase as a reporter. Successful expression of luciferase in B. bovis parasites using this plasmid introduced by classic electroporation of B. bovis infected erythrocytes was followed by the identification and characterization of stronger promoters, such as the ef-1alpha promoter, using transient transfection techniques. Further refinement of the transient transfection technique included development of the ability to transfect free merozoites using nucleofection, an alternative method to electroporation that results in higher transfection yields and improved viability of transfected parasites. Availability of the transient transfection syste

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Author

Carlos E. Suarez; Terry F. McElwain

Publisher

Elsevier Science; Elsevier ; Elsevier BV (ISSN 0304-4017)

Published

2010

Language

EN

Field

Immunology and Microbiology (Life Sciences)