Transient expression directed by homologous and heterologous promoter and enhancer sequences in fish cells.
In order to construct fish specific expression vectors for studies on gene regulation in vitro and in vivo a variety of heterologous enhancers and promoters from mammals and from viruses of higher vertebrate cells were tested for expression of the bacterial chloramphenicol acetyl transferase reporter gene in three teleost fish cell lines. Several viral enhancers were found to be constitutively active at high levels. The human metallothionein promoter showed inducible expression in the presence of heavy metal ions. A fish sequence was isolated that can be used as a homologous constitutively active promoter for expression of foreign genes. Using the human growth hormone gene with an active promoter in fish cells for transient expression insufficient splicing and lack of translation were observed, pointing to limitations in the use of heterologous genes in gene transfer experiments. On the contrary, some heterologous promoters and enhancers functioned in fish cells as well as in their cell type of origin, indicating that corresponding transcription factors are sufficiently conserved between fish and human over a period of 900 million years of independent evolution.
Friedenreich
H
H
Genzentrum/Max-Planck-Institut für Biochemie, Martinsried, FRG.
Schartl
M
M
D000818:Animals; D001483:Base Sequence; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003532:Cyprinodontiformes / Q000235:genetics*; D004742:Enhancer Elements, Genetic; D005786:Gene Expression Regulation; D013006:Growth Hormone / Q000235:genetics; D008546:Melanoma, Experimental; D008668:Metallothionein / Q000235:genetics; D008670:Metals / Q000494:pharmacology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D014176:Protein Biosynthesis; D012326:RNA Splicing; D015183:Restriction Mapping; D014157:Transcription Factors; D014162:Transfection; D014407:Tumor Cells, Cultured
D008670:Metals; D014157:Transcription Factors; D013006:Growth Hormone; D008668:Metallothionein; D015500:Chloramphenicol O-Acetyltransferase