[1]王延洲,梁志清,刘晓芳,等.Smad3基因RNAi慢病毒载体的构建与鉴定[J].第三军医大学学报,2009,31(08):701-702.
 WANG Yan-zhou,LIANG Zhi-qing,LIU Xiao-fang,et al.Construction and identification of lentiviral vector encoding shRNA against Smad3[J].J Third Mil Med Univ,2009,31(08):701-702.
点击复制

Smad3基因RNAi慢病毒载体的构建与鉴定(/HTML )
分享到:

《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
31卷
期数:
2009年第08期
页码:
701-702
栏目:
论著
出版日期:
2009-04-30

文章信息/Info

Title:
Construction and identification of lentiviral vector encoding shRNA against Smad3
作者:
王延洲梁志清刘晓芳徐惠成
第三军医大学西南医院妇产科
Author(s):
WANG Yan-zhou LIANG Zhi-qing LIU Xiao-fang XU Hui-cheng
Department of Obstetrics and Gynecology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China
关键词:
RNA干扰Smad3慢病毒
Keywords:
RNA interference Smad3 lentivirus
分类号:
R373;R394-33;R394.3
文献标志码:
A
摘要:
目的    构建Smad3基因RNAi慢病毒载体。    方法    针对已经筛选确定的Smad3基因RNAi有效靶序列,合成靶序列的Oligo DNA,退火形成双链DNA,与经AgeⅠ和EcoRⅠ双酶切后的pGCSIL-GFP载体连接产生GC-shSmad3慢病毒载体,PCR筛选阳性克隆,测序鉴定。用GC-shSmad3、pHelper 1.0 载体和pHelper 2.0载体共转染包装细胞293T细胞,包装产生慢病毒,以293T细胞GFP蛋白的表达水平测定病毒滴度。    结果    PCR 和测序证实,构建出了Smad3 shRNA的慢病毒载体GC-shSmad3。包装慢病毒,浓缩病毒悬液的滴度为3×108 TU/ml。    结论    成功构建Smad3基因RNAi慢病毒载体。
Abstract:
Objective    To construct a lentiviral vector expressing small-hairpin RNA (shRNA) targeting Smad3 gene.     Methods    The targeting sequence of Smad3 gene which can be effectively silenced in RNA inference was confirmed in our previous study. The cDNA containing both sense and antisense Oligo DNA fragments of the targeting sequence was designed, synthesized and cloned into the pGCSIL-GFP vector. The obtained lentiviral vector containing Smad3 shRNA was confirmed by PCR and sequencing. 293T cells were cotransfected with lentiviral vector GC-shSmad3, pHelper 1.0 and pHelper 2.0. The titer of virus was tested according to the expression level of GFP.     Results    PCR and DNA sequencing demonstrated that the constructed lentivirus vector GC-shSmad3 produced Smad3 shRNA. The titer of concentrated virus was 3×108 TU/ml.     Conclusion    The lentivirus RNAi vector targeting Smad3 is constructed successfully.

参考文献/References:

王延洲,梁志清,刘晓芳,等. Smad3基因RNAi慢病毒载体的构建与鉴定[J]. 第三军医大学学报,2009,31(8):701-702.

相似文献/References:

[1]范婷婷,唐良萏.Nek2基因沉默对卵巢癌SKOV3细胞侵袭能力的影响[J].第三军医大学学报,2012,34(15):1514.
 Fan Tingting,Tang Liangdan.Silencing Nek2 via RNAi suppresses invasiveness in ovarian cancer SKOV3 cells[J].J Third Mil Med Univ,2012,34(08):1514.
[2]梁佳,林力,寿铸,等.鼻咽癌畸胎瘤细胞源性生长因子表达及siRNA对HNE-1细胞株增殖的抑制[J].第三军医大学学报,2012,34(15):1572.
 Liang Jia,Lin Li,Shou Zhu,et al.Expression of PCDGF in nasopharyngeal carcinoma and its silencing in cell proliferation[J].J Third Mil Med Univ,2012,34(08):1572.
[3]刘丹,尹东,汤蕾,等.LPS预处理对心肌细胞缺氧/复氧损伤的保护作用[J].第三军医大学学报,2012,34(17):1707.
 Liu Dan,Yin Dong,Tang Lei,et al.Protective effect of lipopolysaccharide pretreatment on cardiomyocytes anoxia/reoxygenation injury[J].J Third Mil Med Univ,2012,34(08):1707.
[4]赵亮,高蕊,刘菁,等.异位表达FOXO4对胃癌细胞凋亡的影响[J].第三军医大学学报,2015,37(18):1830.
 Zhao Liang,Gao Rui,Liu Jing,et al.Effects of ectopic expression of Forkhead box O4 on cell apoptosis in human gastric cancer cells[J].J Third Mil Med Univ,2015,37(08):1830.
[5]黄士隋,史良会,黄广岩.RNA干扰下调LAT1表达对胃腺癌SGC-7901细胞增殖、侵袭、转移及细胞周期的影响[J].第三军医大学学报,2013,35(04):320.
 Huang Shisui,Shi Lianghui,Huang Guangyan.Effect of RNA interference targeting LAT1 on proliferation, migration and invasion of SGC7901 cells[J].J Third Mil Med Univ,2013,35(08):320.
[6]党微旗,唐浩,曹红,等.可调控STAT3干扰载体抑制BIU-87细胞侵袭的体外研究[J].第三军医大学学报,2013,35(05):400.
 Dang Weiqi,Tang Hao,Cao Hong,et al.Effect of CRE-dependent RNA interference targeting STAT3 on invasion and migration in human bladder cancer BIU-87 cells[J].J Third Mil Med Univ,2013,35(08):400.
[7]张彦,郑晓东,唐鹏,等.Sphk1基因对人乳腺癌MCF-7细胞增殖、凋亡和迁移能力的影响[J].第三军医大学学报,2012,34(21):2141.
 Zhang Yan,Zheng Xiaodong,Tang Peng,et al.Sphk1 interference suppresses proliferation, apoptosis and migration in human MCF-7 breast cancer cells[J].J Third Mil Med Univ,2012,34(08):2141.
[8]王勇,唐川,兰曦,等.沉默CD133基因对CD133+肝癌干细胞放射敏感性的影响[J].第三军医大学学报,2012,34(23):2373.
 Wang Yong,Tang Chuan,Lan Xi,et al.RNAi targeting CD133 enhances radiosensitivity in CD133 positive liver cancer stem cells[J].J Third Mil Med Univ,2012,34(08):2373.
[9]赵亮,李静,魏强,等.人PGI基因siRNA慢病毒质粒的构建及对白血病细胞增殖的影响[J].第三军医大学学报,2013,35(01):20.
 Zhao Liang,Li Jing,Wei Qiang,et al.Construction of a lentiviral vector expressing human phosphoglucose isomerase gene siRNA and its influence on leukemia cell proliferation[J].J Third Mil Med Univ,2013,35(08):20.
[10]熊震,汤旭东,房殿春,等.人肝素酶RNAi序列的筛选及鉴定[J].第三军医大学学报,2007,29(20):1929.
 XIONG Zhen,TANG Xu-dong,FANG Dian-chun,et al.Screening and identification of heparanase RNAi sequence[J].J Third Mil Med Univ,2007,29(08):1929.
[11]但洋,沈为民,果磊.shRNA抑制Smad3基因的表达对瘢痕疙瘩成纤维细胞Smad7表达的影响[J].第三军医大学学报,2009,31(12):1172.
 DAN Yang,SHEN Wei-min,GUO Lei.Effects of the inhibition of Smad3 by short hairpin RNA on the expression of Smad7 in keloid fibroblasts[J].J Third Mil Med Univ,2009,31(08):1172.

更新日期/Last Update: 2009-04-15