Regulation of expression of transgenes in developing fish.
The transcriptional regulatory elements of the beta-actin gene of carp (Cyprinus carpio) have been examined in zebrafish and goldfish harbouring transgenes. The high sequence conservation of the putative regulatory elements in the beta-actin genes of animals suggested that their function would be conserved, so that transgenic constructs with the same transcriptional control elements would promote similar levels of transgene expression in different species of transgenic animals. To test this assumption, we analysed the temporal expression of a reporter gene under the control of transcriptional control sequences from the carp beta-actin gene in zebrafish (Brachydanio rerio) and goldfish (Carrasius auratus). Our results indicated that, contrary to expectations, combinations of different transcriptional control elements affected the level, duration, and onset of gene expression differently in developing zebrafish and goldfish. The major differences in expression of beta-actin/CAT (chloramphenicol acetyltransferase) constructs in zebrafish and goldfish were: (1) overall expression was almost 100-fold higher in goldfish than in zebrafish embryos, (2) the first intron had an enhancing effect on gene expression in zebrafish but not in goldfish, and (3) the serum-responsive/CArG-containing regulatory element in the proximal promoter was not always required for maximal CAT activity in goldfish, but was required in zebrafish. These results suggest that in the zebrafish, but not in the goldfish, there may be interactions between motifs in the proximal promoter and the first intron which appear to be required for maximal enhancement of transcription.
Moav
B
B
Department of Zoology, Tel Aviv University, Israel.
Liu
Z
Z
Caldovic
L D
LD
Gross
M L
ML
Faras
A J
AJ
Hackett
P B
PB
D000199:Actins / Q000096:biosynthesis / Q000235:genetics*; D000818:Animals; D030801:Animals, Genetically Modified; D001483:Base Sequence; D002347:Carps / Q000196:embryology / Q000235:genetics*; D005314:Embryonic and Fetal Development / Q000235:genetics; D005786:Gene Expression Regulation; D005813:Genes, Synthetic; D005822:Genetic Vectors; D006054:Goldfish / Q000196:embryology / Q000235:genetics*; D008845:Microinjections; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid; D013045:Species Specificity; D015027:Zebrafish / Q000196:embryology / Q000235:genetics*
D000199:Actins; D011993:Recombinant Fusion Proteins