Yolk polypeptide gene expression in cultured Drosophila cells.
The transfer of chimaeric plasmids to Drosophila melanogaster cell lines has been examined as a system for investigation of the hormonal regulation of the genes coding for D. melanogaster yolk polypeptide 1 (YP1) and Locusta migratoria vitellogenin B (VgB). Constructs containing promoters and putative 5'-regulatory sequences from these genes, ligated to bacterial chloramphenicol acetyltransferase (CAT) coding DNA, were transfected into Drosophila Kc (Kc-H) and S3 cells, and transient expression of CAT was assayed. Activity was expressed both from the homologous promoter of pYP1CAT and from the heterologous locust promoter of pVgCAT at comparable levels. In S3 cells, with calcium phosphate-mediated transfer of pYP1CAT there was a twofold induction of CAT activity after the addition of 10(-6) M ecdysterone, but no hormonal stimulation was noted when the polycation polybrene was used to achieve transfection. For Kc cells, calcium phosphate was ineffective for transfection, and after transfection with polybrene neither pYP1CAT nor pVgCAT was induced by the juvenile hormone (JH) analog methoprene. It is concluded that S3 cells may be useful for investigating the molecular basis of gene regulation by ecdysteroids, but conditions suitable for the analysis of JH action have not yet been established.
Walker
V K
VK
Department of Biology, Queen's University, Kingston, Ontario, Canada.
Schreiber
M
M
Purvis
C
C
George
J
J
Wyatt
G R
GR
Bendena
W G
WG
D000818:Animals; D002130:Calcium Phosphates / Q000494:pharmacology; D002460:Cell Line; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000502:physiology; D004247:DNA / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics / Q000378:metabolism*; D004441:Ecdysterone / Q000502:physiology; D004527:Egg Proteins / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression / Q000187:drug effects; D006583:Hexadimethrine Bromide / Q000494:pharmacology; D008726:Methoprene / Q000494:pharmacology; D010957:Plasmids / Q000235:genetics / Q000502:physiology; D011401:Promoter Regions, Genetic / Q000235:genetics; D014162:Transfection / Q000235:genetics; D014819:Vitellogenins / Q000235:genetics / Q000378:metabolism / Q000502:physiology
D002130:Calcium Phosphates; D004527:Egg Proteins; D014819:Vitellogenins; C485828:alpha-tricalcium phosphate; C485830:tetracalcium phosphate; D006583:Hexadimethrine Bromide; D004441:Ecdysterone; C485838:calcium phosphate, monobasic, anhydrous; D008726:Methoprene; D004247:DNA; C020243:calcium phosphate; D015500:Chloramphenicol O-Acetyltransferase; C485829:calcium phosphate, dibasic, anhydrous