摘要
为建立硫化物醌氧化还原酶(SQR)的肠道定向表达载体,同时通过转染小鼠肠道上皮细胞确定载体的有效性,本试验采用重叠PCR、中间载体法、酶切连接法获得重组载体,通过脂质体转染法将重组载体转入小鼠肠道上皮细胞,并鉴定重组载体的特异性表达。结果显示,成功构建了以肠脂肪酸结合蛋白质(IFABP)为启动子的SQR肠道定向表达载体pc DNA3.1(-)-IFABP-SQR-GFP,并在小鼠肠道上皮细胞中检测到表达的融合绿色荧光蛋白质(GFP)。表明IFABP启动子能够在肠道细胞中定向启动SQR的表达。
The directed expression vector pc DNA3. 1（-）-IFABP-SQR-GFP for sulfide-quinone reductase（ SQR）gene were successfully constructed by means of nested PCR,intermediate vector and restriction enzyme ligation. The vector was transfected into mouse intestinal epithelial cells by lipofection for the expression of SQR gene. The results revealed that the expression of green fluorescent protein was detected in mouse intestinal epithelial cells,indicative of the expression of SQR gene initiated by the promotor of intestinal fatty acid-binding protein（ IFABP）.
出处
《江苏农业学报》
CSCD
北大核心
2015年第1期100-105,共6页
Jiangsu Journal of Agricultural Sciences
基金
农业部转基因生物新品种培育重大专项(2014ZX08006-004)
江苏省农业科技自主创新基金项目[CX(10)421]
关键词
硫化物醌氧化还原酶
定向表达载体
肠脂肪酸结合蛋白质
肠上皮细胞
sulfide-quinone reductase（SQR）
directed expression vector
intestinal fatty acid-binding protein（IFABP）
intestinal epithelial cell
