Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Chinese Journal of Cellular and Molecular Immunology》 2014-01
Add to Favorite Get Latest Update

Construction of miR-508-5P lentiviral vector and its targeted effect on S-phase kinase-associated protein-2 gene

BAI Jing;XING Yiwen;FAN Weining;GUO Le;LIU Xin;ZHENG Ximing;XU Guangxian;Institute of Laboratory Medicine,Ningxia Medical University;Medical Experimental Center,General Hospital,Ningxia Medical University;  
Objective To construct a lentiviral vector overexpressing miR-508-5p and verify its targeted regulating effect on S-phase kinase-associated protein-2( SKP2) gene. Methods The stem-loop structure RNA of miR-508-5p was obtained by chemical synthesis and cloned into the linearized pSicoR plasmid. The positive recombinants,which had been identified by restriction enzyme digestion and DNA sequencing method,were transfected into HEK293T cells. Meanwhile,the potential target gene of SKP2-3'UTR,which was complementary with miR-508-5p,was obtained and cloned into the linearized pMIRReport plasmid. The targeted effect of miR-508-5p on SKP2 gene was verified by the relative luciferase activity measurement, Western blotting and real-time PCR. Results The PCR,restriction enzyme digestion and DNA sequencing analysis demonstrated that the recombinant plasmids of pSicoR-miR-508-5p and pMIR-Report-SKP2 3'-UTR were constructed successfully. And it was showed that over-expression of miR-508-5p suppressed the mRNA and protein expression level of SKP2 significantly( P 0. 05); and suppression of miR-508-5p expression increased the mRNA and protein expression level of SKP2 significantly( P 0. 05). Conclusion We have successfully constructed the lentiviral vector containing miR-508-5p gene. MiR-508-5p can suppress SKP2 gene expression by targeting the specific sequence of SKP2-3'UTR.
【Fund】: 国家自然科学基金(31160494);; 教育部“新世纪优秀人才支持计划”(NCET 11-11023);; 中国博士后特别基金(2012T50834);; 中国博士后基金(20110491554)
【CateGory Index】: R346
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
Chinese Journal Full-text Database 1 Hits
1 TANG Wei-jun*,ZHANG Xing-ping,ZOU Hong-yuan,PENG Chun-fang(*Department of Oncology,Central Hospital of Jiangjin District,Chongqing City,Chongqing 402260,China);Effect of down-regulation of Skp2 gene expression on biological characters of HepG2 cells as well as expressions of SP1 and p27 proteins[J];Chinese Journal of Biologicals;2012-01
Chinese Journal Full-text Database 10 Hits
1 Yan-chang Sun;Cheng-cheng Guo;Ke Sai;Jian Wang;Jie Wang;Fu-rong Chen;Zong-ping Zhang;Zhong-ping Chen;Guangdong Medical College;Department of Neurosurgery ,Guangdong Medical University Zhanjiang Cancer Hospital;Department of Neurosurgery, Sun Yat-sen University Cancer Center,Guangzhou/State Key Laboratory of Oncology in South China;;miR-181b Enhances Sensitivity to Teniposide in Glioma Cell Line U87[J];Chinese Journal of Neuro-Oncology;2013-02
2 WANG Zhang - xun;ZHOU Quan;MENG Hui - ming;ZHANG Jun;HUANG Bo;School of Plant Protection,Anhui Agricultural University;Anhui Provincial Key Laboratory of Microbial Pest Control,Anhui Agricultural University;;Differential Expression Analysis of MicroRNA in Cicada Slough and Hemolymph Induction Conditions from Metarhizium anisopliae[J];Journal of Anhui Science and Technology University;2013-05
3 Wenwen Jia;Wen Chen;Jiuhong Kang;Clinical and Translational Research Center of Shanghai First Maternity and Infant Health Hospital,Shanghai Key Laboratory of Signaling and Disease Research,School of Life Science and Technology,Tongji University;;The Functions of MicroRNAs and Long Non-coding RNAs in Embryonic and Induced Pluripotent Stem Cells[J];基因组蛋白质组与生物信息学报(英文版);2013-05
4 Chen Juanjuan;Ai Jiangang;Huang Shifeng;Cao Ju;Zhang Liping;Department of Clinical Laboratory,First Affiliated Hospital,Chongqing Medical University;;HBx protein-induced up-regulation of miR-221 promotes HepG2 cell malignant proliferation by targeting ERα[J];Journal of Third Military Medical University;2013-20
5 ZHANG Shi-fang;WEI Cai-hong;LU Jian;ZHANG Xiao-ning;ZHOU Xin-lei;ZHANG Shu-zhen;WANG Guang-kai;CAO Jia-xue;ZHAO Fu-ping;ZHANG Li;DU Li-xin;National Center for Molecular Genetics and Breeding of Animal,Institute of Animal Sciences, Chinese Academy of Agricultural Sciences;;Identification of the microRNAome in Texel Sheep by Deep Sequencing[J];China Animal Husbandry & Veterinary Medicine;2013-09
6 Juan Liu;Chang-Ping Wu;Bin-Feng Lu;Jing-Ting Jiang;Department of Tumor Biological Treatment,The Third Affiliated Hospital of Soochow University;Department of Immunology,University of Pittsburgh School of Medicine;;Mechanism of T cell regulation by microRNAs[J];癌症生物学与医学(英文版);2013-03
7 XIE Xing-yi;LI Yu-guang;HAN Ze-ping;Department of Laboratory,Central Hospital of Panyu District;;Effect of imatinib combined with hsa-miR-203 gene on chronic myeloid leukemia cell line K562[J];Anhui Medical and Pharmaceutical Journal;2013-10
8 CHEN Ling;SHI Chun-mei;YANG Lei;ZHU Lu;XU Guang-feng;JI Chen-bo;GUO Xi-rong;ZHANG Chun-mei;Institute of Pediatrics,Nanjing Medical University;Nanjing Maternal and Child Health Hospital Affiliated to Nanjing Medical University;;Construction of a recombinant lentiviral vector carrying miR-146b and validation of its transduction efficiency in human pre-adipocytes[J];Chinese Journal of Child Health Care;2013-11
9 Mahmoud ElHefnawi;Bangli Soliman;Nourhan Abu-Shahba;Marwa Amer;Centre of Excellence for Advanced Sciences,Informatics and Systems Department,National Research Centre;Stem Cells Research Group,Centre of Excellence for Advanced Sciences,Medical Molecular Genetics Department,National Research Centre;Biology Department,American University in Cairo (AUC);Faculty of Biotechnology,Misr University for Science and Technology (MUST);;An Integrative Meta-analysis of MicroRNAs in Hepatocellular Carcinoma[J];基因组蛋白质组与生物信息学报(英文版);2013-06
10 LI He-wen;WANG Jia-feng;YU Guang;Department of Nephrology,Changhai Hospital,Second Military Medical University;Department of Anesthesiology,Changhai Hospital,Second Military Medical University;;Role of microRNAs in renal ischemia-reperfusion injury:an update[J];Academic Journal of Second Military Medical University;2013-12
China Proceedings of conference Full-text Database 1 Hits
1 Ruiwen Fan Shanshan Yang Zhanquan Shi Kaiyuan Ji Xiaoyun Jia Jianbo Yao George W Smith Changsheng Dong~*;Identification of a novel miRNA important for melanogenesis in alpaca(Lama Pacos)[A];[C];2013
【Secondary Citations】
Chinese Journal Full-text Database 4 Hits
1 LUO Xin,LIU Dan-dan,QI Hong-bo,YAO Zhen-wei,CHEN Guo-qing(Department of Obstetrics and Gynecology,First Affiliated Hospital,Chongqing Medical University,Chongqing 400016,China);Effect of shRNA Lentiviral Vector Targeting Human Growth Arrest and DNA Damage 45α Gene on Disfunction of Human Umbilical Vein Endothelial Cells Due to Hypoxia Stress[J];Chinese Journal of Biologicals;2010-11
2 FANG Li-jun,HUANG Shi-wei,WANG Ying,LIU Ye,ZHANG Shou-feng,HU Rong-liang(College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China);Construction and Identification of Recombinant Lentiviral Vector with Rabies Virus Glycoprotein Gene[J];Chinese Journal of Biologicals;2011-02
3 CHEN Guo-qing,YAO Zhen-wei,QIN Xing-fa(Department of Obstetrics and Gynecology,The First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China);Construction and Identification of Lentivirus-mediated Short Hairpin RNA Targeting Human FOXM1 Gene[J];Chinese Journal of Biologicals;2011-09
4 BAI Xue,DENG Hong(Department of Pathology,College of Medicine,Zhejiang University,Hangzhou 310058,China);Research progress on relationship between transcription factor Sp1 and tumor[J];Journal of Zhejiang University(Medical Sciences);2010-02
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved