Tissue specific expression of antifreeze protein and growth hormone transgenes driven by the ocean pout (Macrozoarces americanus) antifreeze protein OP5a gene promoter in Atlantic salmon (Salmo salar).
Previous research aimed at producing genetically improved salmon broodstock for aquaculture led to the creation of two lines of transgenic Atlantic salmon using gene constructs that were derived in part from the ocean pout OP5a antifreeze protein (AFP) gene. One of the lines was produced using an OP5a AFP gene in which the 5' region of the promoter was removed (termed t-OP5a-AFP), and the other line contains a growth hormone (GH) transgene (EO-1alpha) that consists of a chinook salmon GH cDNA driven by a truncated OP5a AFP promoter that is almost identical to that of the t-OP5a-AFP construct. The similarity of the promoter regions of these transgenes provided an opportunity to evaluate their tissue specific expression patterns. Expression of mRNA was evaluated using Northern blot and RT-PCR techniques. The results demonstrate that the AFP and GH trangenes were expressed in almost all body tissues, suggesting that the promoter region of the OP5a AFP gene lacks tissue specific elements. Northern analysis revealed that expression of the t-OP5a-AFP gene was considerably greater than that of the EO-1alpha GH transgene. Only the spleen tissue of the GH transgenics showed a visible band of hybridization. In contrast clear bands of hybridization were evident in all tissues, except for blood cells, of the AFP transgenics with heart, liver and brain tissue showing the highest levels of mRNA expression. This higher level of expression could be attributable to the presence of introns in the t-OP5a-AFP transgene. Since the GH transgenic salmon grow considerably faster than non-transgenics the low levels of GH transgene expression in this line were clearly sufficient to produce the desired rapid growth phenotype. In contrast the levels of AFP expression were inadequate to impart any improvement in the freeze resistance of the AFP transgenic salmon.
Hobbs
Rod S
RS
Ocean Sciences Centre, Memorial University of Newfoundland, A1C 5S7, St. John's, NL, Canada.
Fletcher
Garth L
GL
D000818:Animals; D030801:Animals, Genetically Modified; D021343:Antifreeze Proteins, Type III / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D005260:Female; D015870:Gene Expression; D013006:Growth Hormone / Q000235:genetics*; D008969:Molecular Sequence Data; D010473:Perciformes / Q000235:genetics*; D011401:Promoter Regions, Genetic; D020317:Salmo salar / Q000235:genetics* / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid; D014018:Tissue Distribution
D021343:Antifreeze Proteins, Type III; D017931:DNA Primers; D018076:DNA, Complementary; D013006:Growth Hormone