摘要
采用lipofectin介导基因转移技术,将含有大肠杆菌β-半乳糖苷酶的结构基因(LacZ基因)及Rous肉瘤病毒启动子的质粒(pRSV-LacZ)导入体外培养的大鼠原代骨骼肌内;另外,分别将具有RSV启动子和巨细胞病毒CMV启动子的LacZ基因直接注射到活体小鼠肌肉组织内。经转染的体外培养骨胳肌细胞和取自活体的肌肉组织切片行X-gal组织化学检测以判断LacZ基因的表达情况。实验结果证明,用lipofectin介导的骨胳肌基因转染率约为10%~20%。直接肌肉注射含有不同启动子的LacZ基因后,肌细胞内均有该基因表达。结果表明,无论在体内或体外,骨骼肌细胞均能被外源性基因所转染并能表达该报告基因。因此,有望使用经遗传工程改造的骨胳肌细胞作为体细胞基因治疗的有效运载细胞。
The plasmid pRSV-LacZ containing report gene─β-galactosidase gene and promoter RSV,was transfected into the primary skeletal muscle cells of rat in vitro by incubating with lipofectin.Meanwhile,the two plasmids,either pRSV-LacZ and pCMV-LacZ,were injected derectly into the M tibialis anterior in mice.The expression of LacZ gene either in the cultrue cells and in the muscle cells was detected respectively by X-gal histochemistry.About 10%~20% of the cultrued primary muscle cells of rat were blue-stained, while a few cells in the sections of M.tibialis anterior in mice were X-gal positive.The results showed that the skeletal muscle cells might be good vehicle for carrying and expressing foreign recommbinant DNA and be useful for somatic gene therapy.
关键词
LACZ基因
骨骼肌
基因表达
DNA
LacZ gene
skeletal muscle
gene expression
DNA transfection