Detection of supercoiled plasmid DNA and luciferase expression in Atlantic salmon (Salmo salar L.) 535days after injection

Specific uptake of plasmid DNA without reporter gene expression in Atlantic salmon (Salmo salar L.) kidney after intramuscular administration

“…Tissue DNA Kit II (Omega Bio-tek, Doraville, GA, USA) according to the manufacturer's recommendation. Total RNA was extracted using the TRIzol method as described earlier [16]. Purified RNA was confirmed to be intact by gel electrophoresis, DNA and RNA concentration and purity was measured spectrophotometrically (NanoDrop Technologies).…”

“…Real-time RTePCR was performed in duplicates with an ABI PRISM â 7000 Sequence Detection System (Applied Biosystems) as described earlier [16]. The luciferase gene was detected using TaqMan probes (Table 1).…”

“…However, a recent report showed intact pDNA in several tissues of Atlantic salmon after i.m. administration [16]. It is not fully understood how the different administration routes influence the distribution of DNA in fish, but it is believed that DNA is transported in blood and, hence, to various organs and tissues after i.m.…”

“…and intravenous (i.v.) administration [5,11,16]. Previous experiments have revealed that the Atlantic salmon kidney has been the major tissue for uptake of DNA fragments after i.v.…”

Development of a suicidal DNA vaccine for infectious hematopoietic necrosis virus (IHNV)

“…Persistence of host cells with luciferase gene constructs has been demonstrated up to two years following vaccination in glass catfish [26]. In another study, supercoiled (intact) plasmid DNA was found at the injection site in Atlantic salmon 535 days post-injection [27]. In rainbow trout, we previously demonstrated that the majority of the injected DNA vaccine remained in muscle cells along the injection path in an unintegrated, unreplicated, lineal or circular conformation [2].…”