Efficient functioning of plant promoters and poly(A) sites in Xenopus oocytes.
Mature Xenopus oocytes were challenged with DNA constructs including plant regulatory elements, namely, the Cauliflower mosaic virus (CaMV) 35S promoter as well as the nopaline synthase (NOS) promoter and polyadenylation signal. The bacterial chloramphenicol acetyl transferase (CAT) was used as a reporter gene. When microinjected into these cells, the plant-derived DNA constructs effectively promoted CAT synthesis in a manner dependent on the presence of the plant promoters and probably also on the polyadenylation signals. Structural studies revealed that the supercoiled structures of the above DNA plasmids were much more active in supporting CAT synthesis in microinjected oocytes than their linear forms, with clear correlation between efficient gene expression and DNA topology. In contrast, the linear forms of these plasmids were considerably more active than the supercoiled ones in transfected plant protoplasts. These findings demonstrate, for the first time, the activity of regulatory elements from plant genes in Xenopus oocytes and shed new light on the specific rules applicable for gene expression in plant and animal cells.
Ballas
N
N
Department of Biological Chemistry, Hebrew University, Jerusalem, Israel.
Broido
S
S
Soreq
H
H
Loyter
A
A
D000594:Amino Acid Oxidoreductases / Q000235:genetics; D000818:Animals; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000235:genetics*; D005260:Female; D008845:Microinjections; D009029:Mosaic Viruses / Q000235:genetics; D009865:Oocytes / Q000378:metabolism*; D010944:Plants / Q000235:genetics*; D010957:Plasmids; D011061:Poly A / Q000235:genetics*; D011401:Promoter Regions, Genetic; D012045:Regulatory Sequences, Nucleic Acid; D014162:Transfection; D014981:Xenopus
D011061:Poly A; D004247:DNA; D000594:Amino Acid Oxidoreductases; C021883:nopaline synthase; D015500:Chloramphenicol O-Acetyltransferase