质粒DNA在大鼠舌肌中的表达  

Plasmid DNA Expression in the Muscle of Rat Tongue

作　　者：高文涛[1] 张泽兵[1] 欧阳喈[1] 

机构地区：[1]白求恩医科大学口腔医学院,130041

出　　处：《华西口腔医学杂志》1999年第4期297-299,共3页West China Journal of Stomatology

基　　金：卫生部基金!资助项目 (编号 9412 17)

摘　　要：目的:建立质粒DNA大鼠舌肌表达模型。方法:将表达质粒pM3TnC3、pSV40-LacZ直接注射入Wistar大鼠舌肌中,应用PCR技术、β-半乳糖苷酶(X-gal)组织化学染色及Southern杂交技术分析质粒携带报导基因——乳糖苷酶(LacZ)体内表达情况。结果:PCR结果表明大鼠舌肌纤维具有摄取外源DNA的能力。报导基因表达与质粒DNA注射量及在舌肌中孵化时间相关。注射后24h可检测出大肠杆菌β-半乳糖苷酶的活性,2个月时仍可检测到。但表达水平在1周时达到高峰。舌肌中共同注射两种表达质粒可在同一横纹肌纤维中同时表达。Southern杂交显示质粒DNA游离于舌肌细胞基因组之外,未发生整合。结论:舌肌中直接注射质粒DNA是一种简便有效的将外源基因转入横纹肌细胞中的方法。Objective: To develop rat tongue model for plasmid DNA expression in vivo. Methods: The expression vectors which contained either human muscle specific promoter or SV40 promoter directing the transcription of reporter gene (E.coli LacZ gene) were used. The striated muscle of the tongue was used as a model system, and expression plasmid DNA was injected directly into Wistar rat tongue. PCR technique, X gal histochemical staining and Southen blot hybridization were used to analyze reporter gene expression in vivo. Results: The expression of reporter gene was correlated with the amount of injected DNA and the time of incubation in tongue muscle. β galactosidase activity can be detected 24 hours after injection of X gal staining and even be detected after two months, but the maximal expression level was observed one week later. Coinjection of two expression plasmids resulted in co expression in the same striated myofibers. E. coli LacZ DNA sequence was found in rat tongue injected with the plasmid DNA by PCR, which demonstrated capacity of the rat tongue myofibers uptaking foreign DNA. The Southern blot analysis showed the injected plasmid DNA existed outside the genome of rat muscle cells. No integration was detected. Conclusion: Direct plasmid DNA injection into tongue muscle was a simple and efficient approach to transfer foreign gene into striated muscle cells, and tongue was a good model for analyzing exogenous gene expression.\;

关 键 词：质粒 报导基因 舌肌 大鼠 DNA 舌癌 

分 类 号：R739.860.2[医药卫生—肿瘤]