Biolistic transformation of Schistosoma mansoni: Studies with modified reporter-gene constructs containing regulatory regions of protease genes

0352015 - BC 2011 RIV NL eng J - Journal Article
Dvořák, Jan - Beckmann, S. - Lim, K.-C. - Engel, J. C. - Grevelding, C. G. - McKerrow, J. H. - Caffrey, C. R.
Biolistic transformation of Schistosoma mansoni: Studies with modified reporter-gene constructs containing regulatory regions of protease genes.
Molecular and Biochemical Parasitology. Roč. 170, č. 1 (2010), s. 37-40. ISSN 0166-6851. E-ISSN 1872-9428
Institutional research plan: CEZ:AV0Z60220518
Keywords : Schistosoma * Protease * Transgene * Gene promoter * Biolistics * Electroporation
Subject RIV: EB - Genetics ; Molecular Biology
Impact factor: 2.875, year: 2010

Biolistics of the flatworm parasite Schistosoma mansoni facilitates the accurate spatial expression of transgenes under the control of gene-specific promoter elements. To improve transgene expression, either in the number of positive worms and/or an increased transgene signal per worm, we tested plasmid constructs incorporating 5_ and 3_ gene-specific genomic fragments, and parts of the open reading frame for two S. mansoni proteases, cathepsins F and D (SmCF and SmCD). GFP-expression was gut-localized, a novel finding for SmCD and consistent with previous data for SmCF. The mCherry fluorescent protein can also operate as a reporter. Though certain constructs imparted stronger and better distributed signals per positive worm, the low yields throughout precluded further quantifications of improvement. Electroporation of the same constructs was also weakly efficient. However, reporter signals were found in tissues other than the gut, which may represent dysregulated transcription.
Permanent Link: http://hdl.handle.net/11104/0191626