Transfer of a functional arabinose operator-repressor system to Drosophila melanogaster Schneider line-2 cells.
Regulatory elements from the arabinose operon of Escherichia coli were transfected into Drosophila melanogaster Schneider line 2 cells to test their ability to function in animal cells. A construct containing an araC fusion gene (encoding AraC protein) under the control of an act5C (encoding actin) promoter and a construct containing a lacZ fusion gene (reporter gene) also under the control of an act5C promoter were used to build a regulatory circuit in the D. melanogaster cells. We have demonstrated that a AraC fusion protein can be synthesised in Schneider cells where it is able to repress the transcription of the reporter gene containing AraC-binding sites inserted between the transcription start point and the initiation codon. The reporter gene activity can be further modulated by the addition of arabinose to the medium.
Raman
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CSIRO Division of Biomolecular Engineering, North Ryde, NSW, Australia.
Rand
K N
KN
Hill
R J
RJ
D000818:Animals; D051777:AraC Transcription Factor; D001089:Arabinose / Q000494:pharmacology; D001426:Bacterial Proteins; D001483:Base Sequence; D002467:Cell Nucleus / Q000378:metabolism; D017931:DNA Primers / Q000737:chemistry; D004331:Drosophila melanogaster / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics*; D029968:Escherichia coli Proteins; D005786:Gene Expression Regulation; D015964:Gene Expression Regulation, Bacterial; D008969:Molecular Sequence Data; D009875:Operator Regions, Genetic; D009876:Operon; D012097:Repressor Proteins / Q000502:physiology*; D014157:Transcription Factors
D051777:AraC Transcription Factor; C497371:AraC protein, E coli; D001426:Bacterial Proteins; D017931:DNA Primers; D029968:Escherichia coli Proteins; D012097:Repressor Proteins; D014157:Transcription Factors; D001089:Arabinose