Gene expression in rainbow trout (Oncorhynchus mykiss) following intramuscular injection of DNA.

E D Anderson, D V Mourich, J A Leong
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Abstract

Expression of the firefly luciferase gene under the control of viral or fish promoters was observed in fish tissue after direct DNA injection of plasmid DNA. Plasmid DNA containing the firefly luciferase gene was injected into the skeletal muscle of rainbow trout (Oncorhynchus mykiss), and levels of luciferase activity were found to be dependent on the controlling promoter and the amount of injected DNA. Plasmids using the cytomegalovirus immediate early promoter (CMV-IEP) consistently produced the highest levels of luciferase activity. Maximal activity was observed five to seven days postinjection with 50 micrograms of DNA. This activity persisted in the tissues for as long as 115 days postinjection. When the DNA was examined up to two months postinjection, the predominant form was unreplicated, unintegrated DNA in linear and relaxed circular conformation. Expression of injected DNA was found predominantly within muscle cells along the injection path and in scattered muscle cells anterior to the injection site.

虹鳟鱼肌肉注射DNA后基因表达。
直接注射质粒DNA后,在病毒或鱼类启动子的控制下,在鱼组织中观察到萤火虫荧光素酶基因的表达。将含有萤火虫荧光素酶基因的质粒DNA注入虹鳟鱼(Oncorhynchus mykiss)的骨骼肌,发现荧光素酶活性水平依赖于控制启动子和注射DNA的量。使用巨细胞病毒即时早期启动子(CMV-IEP)的质粒始终产生最高水平的荧光素酶活性。注射50微克DNA后5至7天观察到最大活性。这种活性在注射后的组织中持续了115天。当DNA在注射后两个月检查时,主要形式是线性和松弛圆形构象的未复制,未整合的DNA。注射DNA的表达主要在沿注射路径的肌肉细胞和注射部位前的分散肌肉细胞中发现。
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