Cloning and characterization of Echinococcus granulosus (Cestode) EgactI and EgactII actin gene promoters and their functional analysis in the NIH3T3 mouse cell line.
We report here for the first time the structure and function of a promoter from a cestode. The ability of DNA fragments respectively encompassing the 935-bp and 524-bp regions upstream from the ATG codon from the EgactI and EgactII actin genes of Echinococcus granulosus to promote transcription was studied in the NIH3T3 mouse cell line. The results of transfection assays showed that both regions have strong promoter activity in these cells. The fragments were tested in both orientations and the 524-bp fragment of EgactII presented a bidirectional promoter activity. Deletion analysis of EgactI and EgactII promoters indicated the presence of regulatory regions containing putative silencer elements. These results indicate that both EgactI and EgactII promoters are functional and that the preliminary functional evaluation of E. granulosus and possibly of other cestode promoters can be performed in heterologous cell lines.
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Departamento de Biologia Molecular e Biotecnologia and Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brasil. etel@bioqmed.ufrj.br
Chemale
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Farias
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Zaha
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D016475:3T3 Cells / Q000201:enzymology*; D000199:Actins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000378:metabolism*; D003001:Cloning, Molecular; D004446:Echinococcus / Q000235:genetics*; D015870:Gene Expression; D017930:Genes, Reporter; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D051379:Mice; D011401:Promoter Regions, Genetic / Q000235:genetics / Q000502:physiology*; D013329:Structure-Activity Relationship
C431644:ACTI protein, Echinococcus granulosus; C431645:ACTII protein, Echinococcus granulosus; D000199:Actins; D015801:Helminth Proteins; D015500:Chloramphenicol O-Acetyltransferase