[Transformation of schistosomulae by electroporation and transient expression of the enhanced green fluorescent protein (EGFP) gene].
OBJECTIVE
To explore the possibility of heterogeneous gene to express in juvenile Schistosoma japonicum and the application of electroporation in transformation of schistosomulae.


METHODS
The plasmids of pEGFP-C1 were introduced into mechanically transformed schistosomula with electroporation. The presence, transcription and translation of the transgene in electroporated schistosomula were confirmed by PCR, RT-PCR and Western blotting analysis respectively using the genomic DNA, total RNA and protein extracted and isolated from schistosomula cultured in vitro for 48 hours. Meanwhile, localization of EGFP within electroporated schistosomula was performed with confocal laser scanning micro scope.


RESULTS
760 bp and 276 bp amplified products by PCR and RT-PCR were found coincident with the expected size and expression of EGFP gene in electroporated schistosomula was confirmed by Western blotting. Fluorescence of EGFP was localized in tegument and subtegument of the electroporated schistosomula with confocal microscopy, especially in the anterior part of the worm.


CONCLUSIONS
The heterogeneous gene of EGFP has been successfully introduced into juvenile S. japonicum by electroporation and the expression of transgene was confirmed with molecular and microscopical methods.
Yuan
Xiao-song
XS
Department of Microbiology and Parasitology, Anhui Medical University, Hefei 230032, China.
Shen
Ji-long
JL
Wang
Xue-long
XL
Hu
Yuan-sheng
YS
Luo
Qing-li
QL
D000818:Animals; D015153:Blotting, Western; D018274:Electroporation; D015870:Gene Expression; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012549:Schistosoma japonicum / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins