Tan J H, Chan W K
School of Biological Sciences, National University of Singapore, Republic of Singapore.
Mol Mar Biol Biotechnol. 1997 Jun;6(2):98-109.
The ability of zebrafish skeletal muscles to internalize and express plasmid DNA was demonstrated using pCMVCAT1, a chloramphenicol acetyltransferase (CAT) construct driven by the human cytomegalovirus immediate early (CMV-IE) promoter. We found that CAT activity was correlated to the amount of plasmid DNA injected, with maximal expression at 5 micrograms of pCMVCAT1. CAT activity was also shown to increase steadily over the first seven days after injection, with high levels of CAT expression persisting up to one year. Intramuscular injection of CAT constructs driven by other viral promoters also resulted in high levels of CAT activity. Histochemical localization using a CMV beta-galactosidase construct confirmed that only myofibers at the site of injection expressed beta-galactosidase enzyme. The persistence and strong expression of injected plasmid constructs suggest that zebrafish may be a simple and readily accessible system for direct muscle injection studies.
使用pCMVCAT1证明了斑马鱼骨骼肌内化和表达质粒DNA的能力，pCMVCAT1是一种由人巨细胞病毒立即早期（CMV-IE）启动子驱动的氯霉素乙酰转移酶（CAT）构建体。我们发现CAT活性与注射的质粒DNA量相关，在注射5微克pCMVCAT1时表达最高。还显示CAT活性在注射后的前七天稳步增加，高水平的CAT表达持续长达一年。由其他病毒启动子驱动的CAT构建体的肌肉内注射也导致高水平的CAT活性。使用CMVβ-半乳糖苷酶构建体的组织化学定位证实，只有注射部位的肌纤维表达β-半乳糖苷酶。注射的质粒构建体的持久性和强表达表明，斑马鱼可能是用于直接肌肉注射研究的简单且易于使用的系统。
