Fish transgene expression by direct injection into fish muscle.
The ability of a promoter sequence to drive expression of a reporter gene can be determined by direct injection of copies of the cloned sequence into fish muscle, followed by biopsy of muscle from the site of injection. We describe a set of experiments in which copies of the constructs FV1 and FV2, both comprising a carp beta-actin promoter sequence spliced to the bacterial reporter gene CAT, were injected into the muscle of tilapia fish )Oreochromis niloticus) of between 5 and 8 cm body length. The site of injection was carefully determined so that biopsy samples could be recovered from the injection site 24 hours, 48 hours, and 7 days after injection. Biopsy samples of muscle were homogenized and used for CAT assays. CAT activity was successfully detected in many of the muscle samples.
Rahman
A
A
Department of Biology, Southampton University, Hampshire, UK.
Maclean
N
N
D000199:Actins / Q000235:genetics; D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D001706:Biopsy / Q000662:veterinary; D015500:Chloramphenicol O-Acetyltransferase / Q000096:biosynthesis* / Q000235:genetics; D004247:DNA / Q000008:administration & dosage; D015870:Gene Expression; D007273:Injections, Intramuscular / Q000662:veterinary; D009132:Muscles / Q000201:enzymology*; D011401:Promoter Regions, Genetic; D012588:Scintillation Counting; D017210:Tilapia / Q000235:genetics*
D000199:Actins; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase