Strongyloides stercoralis: cell- and tissue-specific transgene expression and co-transformation with vector constructs incorporating a common multifunctional 3' UTR.
Transgenesis is a valuable methodology for studying gene expression patterns and gene function. It has recently become available for research on some parasitic nematodes, including Strongyloides stercoralis. Previously, we described a vector construct, comprising the promoter and 3' UTR of the S. stercoralis gene Ss era-1 that gives expression of GFP in intestinal cells of developing F1 progeny. In the present study, we identified three new S. stercoralis promoters, which, in combination with the Ss era-1 3' UTR, can drive expression of GFP or the red fluorescent protein, mRFPmars, in tissue-specific fashion. These include Ss act-2, which drives expression in body wall muscle cells, Ss gpa-3, which drives expression in amphidial and phasmidial neurons and Ss rps-21, which drives ubiquitous expression in F1 transformants and in the gonads of microinjected P0 female worms. Concomitant microinjection of vectors containing GFP and mRFPmars gave dually transformed F1 progeny, suggesting that these constructs could be used as co-injection markers for other transgenes of interest. We have developed a vector "toolkit" for S. stercoralis including constructs with the Ss era-1 3' UTR and each of the promoters described above.
Junio
Ariel B
AB
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA.
Li
Xinshe
X
Massey
Holman C
HC
Nolan
Thomas J
TJ
Todd Lamitina
S
S
Sundaram
Meera V
MV
Lok
James B
JB
D020413:3' Untranslated Regions / Q000502:physiology*; D000818:Animals; D017173:Caenorhabditis elegans / Q000235:genetics; D018509:DNA, Helminth / Q000008:administration & dosage; D004285:Dogs; D005260:Female; D005786:Gene Expression Regulation / Q000502:physiology*; D017930:Genes, Reporter / Q000235:genetics; D005822:Genetic Vectors; D005849:Gerbillinae; D049452:Green Fluorescent Proteins / Q000235:genetics; D008845:Microinjections; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000235:genetics; D012692:Serial Passage; D017171:Strongyloides stercoralis / Q000235:genetics*; D014170:Transformation, Genetic / Q000502:physiology*; D019076:Transgenes / Q000502:physiology*
D020413:3' Untranslated Regions; D018509:DNA, Helminth; D049452:Green Fluorescent Proteins