裸DNA骨骼肌注射表达外源基因条件的初步研究

PRELIMINARY STUDY OF THE CONDITIONS OF FOREIGN GENE EXPRESSION BY INTRAMUSCULAR INJECTION OF NAKED DNA

目的观察裸ＤＮＡ直接骨骼肌注入表达外源基因的条件。方法以真核表达质粒为载体将报告基因β－ｇａｌ导入骨骼肌细胞，冰冻切片后Ｘ－ｇａｌ染色检测β－ｇａｌ基因表达水平。结果所测质粒ＤＮＡ溶剂类型中，生理盐水最有利于β－ｇａｌ表达；表达水平与注射ＤＮＡ的含量呈剂量依赖性；注射体积对表达影响不明显。结论ＤＮＡ溶液中的溶质成份影响外源基因在肌肉中的表达，优选这些成份可进一步提高肌注后基因表达水平。

Objective: To observe the conditions of foreign gene expression by direct intramusular injection of naked DNA. Methods: The expression level of β-gal gene which was introduced by eukaryotic plasmid expression vector into skeletal muscles which were cryosctioned after X-gal stainage was identified. Results: Among the different exmined solutions, normal saline was best suited to gene expression; the relation between expression levels and DNA injection dose was in dose-reponse; the effect of injection volume on gene expression levels was not obvious. Conclusion: Foreign gene expression levels in skeletal muscles are affected by the components of DNA solutions, optimization of which helps improve gene expression levels after intramuscular injection.