摘要
试验通过提取绒山羊皮肤总RNA,克隆角蛋白辅助蛋白1.1(keratin-associated protein 1.1,KAP1.1)基因,构建重组真核表达载体pEGFP-N3-KAP1.1,利用脂质体介导法将重组质粒转入内蒙古绒山羊体外培养的成纤维细胞中,研究该基因在真核细胞中的时空表达、亚细胞定位及表达规律。结果显示,2.0μg重组质粒和6.0μL脂质体的比例为适合本试验的最优化方案,KAP1.1基因能在内蒙古绒山羊成纤维细胞中成功表达,并在转染后48h转染率达到峰值,该基因在真核体外培养细胞中细胞核、细胞质和细胞膜上均有表达,但以细胞核表达量最强。
In this study, Cashmere goat skin by extracting total RNA, cloning keratin-associated protein 1.1 （KAP1.1） gene eukaryotic expression vector pEGFP-N3 KAP1.1, using liposome mediated recombinant plasmids were transformed into Inner Mongolia Cashmere goats cultured fibroblasts, temporal and spatial expression of the genes in eukaryotic ceils, subceliu- lar localization and expression pattern. The results showed that 2.0μg plasmid and 6.0μL ratio liposomes suitable for the opti- mization of the experimental protocol, KAP1.1 gene could be successfully expressed in fibroblasts in Inner Mongolia Cashmere goat, and 48 h after transfection, the transfection efficiency reaches a peak, the gene in eukaryotic cells in vitro on the nucleus, cytoplasm and cell membrane are expressed, but the strongest expression of the nucleus.
出处
《中国畜牧兽医》
CAS
北大核心
2014年第3期39-43,共5页
China Animal Husbandry & Veterinary Medicine
基金
内蒙古自治区教育厅高校科研项目自然科学重点项目(NJZZ11277)
内蒙古自然科学基金项目(2013MS0416)
