Anderson E D, Mourich D V, Leong J A
Department of Microbiology, Oregon State University, Corvallis 97331, USA.
Mol Mar Biol Biotechnol. 1996 Jun;5(2):105-13.
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.
在将质粒DNA直接注射到鱼组织后，观察到在病毒或鱼类启动子控制下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注射到虹鳟（Oncorhynchus mykiss）的骨骼肌中，发现荧光素酶活性水平取决于控制启动子和注射的DNA量。使用巨细胞病毒立即早期启动子（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。这种活性在注射后长达115天的时间内持续存在于组织中。当在注射后长达两个月检查DNA时，主要形式是呈线性和松弛环状构象的未复制、未整合的DNA。发现注射的DNA主要在注射路径沿线的肌肉细胞内以及注射部位前方的散在肌肉细胞中表达。
