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| BIRC5 shRNA慢病毒表达载体的构建与鉴定 | |
| 引用本文： | Rao GZ,Li A,Zhu YJ,Shi JF,Gou JZ,Zhang YC. BIRC5 shRNA慢病毒表达载体的构建与鉴定[J]. 细胞与分子免疫学杂志, 2011, 27(7): 767-769 |
| 作者姓名： | Rao GZ Li A Zhu YJ Shi JF Gou JZ Zhang YC |
| 作者单位： | 1. 西安交通大学口腔医院中心实验室,陕西,西安,710004 2. 西安交通大学口腔医院牙周科,陕西,西安,710004 3. 西安交通大学口腔医院颌面外科,陕西,西安,710004 |
| 基金项目： | 陕西省社发攻关资助项目 |
| 摘 要： | 目的:应用RNA干扰技术,以人BIRC5基因为靶基因,设计合成3对短发夹结构的互补DNA序列和一对阴性对照序列,构建慢病毒干扰载体并鉴定。方法:将设计合成的单链引物退火成双链oligo序列,连接入经AgeI和EcoR I双酶切线性化的pMAGic慢病毒质粒载体中,经转化DH5α感受态细胞并筛选出阳性克隆,采用PCR扩增和DNA测序鉴定阳性克隆。结果:PCR扩增产物经凝胶电泳阳性克隆得到335 bp条带,阴性克隆得到298 bp条带,DNA测序结果证实其含设计合成序列。结论:4对BIRC5 shRNA重组慢病毒表达载体构建成功,为研究靶向BIRC5 siRNA对肿瘤细胞增殖抑制与诱导凋亡作用及基因治疗研究奠定了实验基础。 |
| 关 键 词： | RNA干扰 BIRC5 慢病毒表达载体 |
Construction and identification of BIRC5 shRNA lentiviral expression vector |
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| Rao Guo-zhou,Li Ang,Zhu Yong-jin,Shi Jian-feng,Gou Jian-zhong,Zhang Yin-cheng. Construction and identification of BIRC5 shRNA lentiviral expression vector[J]. Chinese journal of cellular and molecular immunology, 2011, 27(7): 767-769 | |
| Authors: | Rao Guo-zhou Li Ang Zhu Yong-jin Shi Jian-feng Gou Jian-zhong Zhang Yin-cheng |
| Affiliation: | RAO Guo-zhou1,LI Ang1,ZHU Yong-jin1,SHI Jian-feng1,GOU Jian-zhong2,ZHANG Yin-cheng31Research Center of Stomatology,2Department of Periodontology,3Department of Oral Maxillofacial Surgery,Stomatological Hospital,Medicine College of Xi'an Jiaotong University,Xi'an 710004,China |
| Abstract: | AIM:Design and synthesis complementary DNA sequences of 3 pairs of short hairpin structure and a pair of negative control sequence by RNA interference technique and construct and identify a lentiviral interference vector with human BIRC5 gene as target gene.METHODS:The designed and synthesised Single-Stranded primer were annealed to Double-Stranded oligo sequences and subcloned into linear pMAGic lentiviral plasmid vector digested by enzyme Age I and EcoR I.Screening positive clone after transformed into DH... |
| Keywords: | BIRC5 |
| 本文献已被 CNKI 万方数据 PubMed 等数据库收录！ | |