脂质体法转染山羊骨骼肌特异细胞条件的优化

Optimization of Liposome Mediated Transfection into Goat Skeletal Muscle Cells

试验旨在通过脂质体转染法检测pIMAU-EGFP载体在山羊骨骼肌细胞中的表达情况，并探索其最佳的转染条件。在构建的适合携带外源基因的山羊骨骼肌特异表达载体pIMAU-EGFP基础上，以绿色荧光蛋白为报告基因，采用脂质体转染法检测了pIMAU-EGFP载体在山羊骨骼肌细胞中的表达，并对转染条件进行了摸索。试验结果表明，成功把测序验证正确的pIMAU-EGFP载体转染到山羊骨骼肌细胞中，筛选的最佳转染条件：脂质体与质粒的最佳比例为1：2，质粒浓度为3.5 μg/μL，细胞密度不低于80%，以转染24-36 h观察记录转染效率为宜。本试验初步建立了转染山羊骨骼肌细胞的方法，绿色荧光蛋白可作为报告基因优化脂质体转染条件，这将为快速和规模化筛选山羊肉质特性相关基因提供新思路和研究方法。

The aim of this study was to screen the optimized transfection conditions for liposome mediated gene transferring into skeletal muscle cells based on the conducted specific expression vector （pIMAU-EGFP）, which was applied to the human gene therapy. The vector could be inserted the ectogenous genes of goats. Green fluorescence protein was conducted as the reporter gene. The results revealed that the optimized transfection conditions for goat skeletal muscle cells were that the ratio of liposome and DNA was 1：2, for 24-36 h and the concentration of DNA was 3.5 μg/μL. Also the cell densities should be over 80%. This experiment established a method to transfect foreign genes into skeletal muscle cells in goat. Green fluorescent protein could be used as the reportor gene. It would provide new ideas and methods for rapid and large-scale screening the quality related genes in goats.