肌肉中特异表达真核载体的构建及表达特异性验证  

Construction and verification of muscle-specific expression eukaryotic vector

作　　者：赵为民[1] 任守文[1] 方晓敏[1] 李碧侠[1] 涂枫[1] 付言峰[1] 赵芳[1] 王学敏[1] 

机构地区：[1]江苏省农业科学院畜牧研究所/动物品种改良和繁育重点实验室,江苏南京210014

出　　处：《江苏农业学报》2016年第6期1359-1366,共8页Jiangsu Journal of Agricultural Sciences

基　　金：转基因生物新品种培育重大专项(2014ZX08006-003);国家生猪现代产业技术体系(CARS-36)

摘　　要：为构建肌肉中特异表达真核载体,本试验以PGL4.10质粒为骨架,利用基因合成、酶切连接方法获得重组载体,同时利用红色荧光蛋白基因DsRed2作为标记基因来检测重组载体在非肌源细胞和肌细胞中的表达情况。结果表明,成功构建了含有肌球蛋白轻链基因MLC启动子及其增强子的肌肉中特异表达载体,通过mRNA和蛋白水平检测,该表达载体在非肌源性细胞和肌细胞中均不表达DsRed2,只在分化的肌细胞中表达DsRed2。该表达载体的构建为制备肌肉中特异表达的转基因动物奠定了基础。To construct a muscle-specific eukaryotic expression vector and to verify its specific expression,a recom-binant vector was constructed using gene synthesis and enzyme digestion ligation method based on PGL4. 10 plasmid as a backbone. Meanwhile,a red fluorescent protein （ DsRed2） was used as a marker to detect the expression of the recombi-nant vector in non-muscle-derived cells and muscle cells. The results showed that the muscle-specific eukaryotic expression vector containing the promoter and enhancer of myosin light chain （ MLC） gene was successfully constructed. This expres-sion vector did not express DsRed2 in non-myogenic cells and muscle cells, and could only express in differentiated muscle cells at mRNA and protein levels. The results provide a solid foundation for the preparation of muscle-specific expression for transgenic animal.

关 键 词：肌肉 表达载体 MLC 启动子 DsRed2 肌细胞 

分 类 号：Q782[生物学—分子生物学]