Full-Text Search:
Home|Journal Papers|About CNKI|User Service|FAQ|Contact Us|中文
《Journal of Northeast Agricultural University》 2011-04
Add to Favorite Get Latest Update

Integrating specific sequence into target gene using PCR method

XU Yaying,WANG Yong,LUO Hong,CHEN Dian(College of Horticulture,Northeast Agricultural University,Harbin 150030,China)  
An intron was introduced into a cre gene by one-step PCR,two-step PCR,three-step PCR amplification strategy,respectively,and using three primers and two templates,in order to achieve the purpose of genetic modification.In the study,it was found that by one-step PCR amplication strategy,target genes were amplified difficultly.By two-step PCR strategy,target genes could be found,but existed non-specific amplification.By three-step PCR strategy,a linear PCR was performed using the intron fragments from the above step as the primer,the resulting products from the second step were served as the template to amplify.Target genes could be amplified efficiently.It concluded that three-step PCR strategy was an ideal method.
【Fund】: 黑龙江省青年基金资助项目(QC07C46);; 东北农业大学博士启动基金资助项目
【CateGory Index】: Q78
Download(CAJ format) Download(PDF format)
CAJViewer7.0 supports all the CNKI file formats; AdobeReader only supports the PDF format.
【Citations】
Chinese Journal Full-text Database 7 Hits
1 Chen Shuangxi et al.(Institute of Bioengineering,College of Life Science,Henan University,Kaifeng 475004,China);Site-specific Recombination Technique Mediated by Cre/LoxP System[J];Anhui Agricultural Science Bulletin;2009-15
2 QIU Shu-ping1,2,CHEN Zai-jie1,WANG Feng1(1.Fujian Key Laboratory of Genetic Engineering for Agriculture,Fujian Academy of Agricul-tural Sciences,Fuzhou,Fujian 350003,China;2.College of Life Sciences,Fujian Normal University,Fuzhou,Fujian 350007,China);Application of Cre/loxp site-specific recombination technology on transgenic plants[J];Fujian Journal of Agricultural Sciences;2008-02
3 Xu Fang1,2 Yao Quanhong2 Xiong Aisheng2 Peng Rihe2 Hou Xilin1* Cao Bing3* 1 College of Horticulture, Nanjing Agricultural University, Nanjing, 210095; 2 Agro-Biotechnology Research Center of Shanghai Academy of Agricultural Sciences, Shanghai, 201106; 3 Sanya Science &Technology Academy for Crop Winter Multiplication, Sanya, 572000;SOE PCR and its Application in Genetic Engineering[J];Molecular Plant Breeding;2006-05
4 ZHAO He(Institute of Genetics and Physiology,Hebei Academy of Agriculture and Forestry Sciences,Shijiazhuang 050051,China);Application of Biotechnology in Mutagenesis[J];Journal of Hebei Agricultural Sciences;2006-01
5 WANG Wen-Qi1), GAI Ying1), LU Hai1), LI Yi2), JIANG Xiang-Ning1) ** (1)College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China; 2)Laboratory of Transgene, The University of Connecticut, Connecticut, Unit 3088 Storrs, USA);Recombinase Cre Mediated DNA Recombination and Gene's Transferring Between Vectors[J];Progress in Biochemistry and Biophysics;2007-11
6 WANG Qiu-ying(Basal Department of Hebei Tourism Vocational College,Chengde Hebei 067000,China);The Technology of Site-directed Mutagenesis[J];Journal of Shanxi Agricultural University;2007-S2
7 Li Daxu Ma Jingyun Cao Yongchang(1College of Animal Science,South China Agricultural University,Guangzhou 510642;2College of Life Science,Sun Yat-Sen University,Guangzhou 510275);Characteristic and Application in Antibody Research of Cre-LoxP Recombination System[J];Biotechnology Bulletin;2008-01
【Co-citations】
Chinese Journal Full-text Database 10 Hits
1 YANG Yang et al(College of Life Sciences,Nanjing Xiaozhuang University,Nanjing,Jiangsu 211171);Research on the Poplar Transferred of AtNHX1 Gene with the cre/lox Expression Vector[J];Journal of Anhui Agricultural Sciences;2009-19
2 ZHU XIN-HONG,GAO MEI-HUA,REN SHU-RONG,et al(Department of Immunology,Qingdao University Medical College,Qingdao 266071,China);SITE-DIRECTED MUTAGENESIS OF CD59 GENE AND CONSTRUCTION OF EUKARYOTIC EXPRESSION SYSTEM[J];Acta Academiae Medicinae Qingdao Universitatis;2008-02
3 FAN Xiang-jun,ZHOU Yuan,ZHU Ming-yan,WANG Zhi-wei Department of General Surgery,Affiliated Hospital of Nantong University,Nantong 226001,China;Transplantation of bone marrow mesenchymal stem cells harboring fusing gene of enhanced green fluorescent protein and human insulin gene in treatment of diabetic mice[J];Academic Journal of Second Military Medical University;2008-12
4 CHEN Guo-liang1,2;ZHANG Jin-wen2,WANG Di2,CHEN Zong-li1,YANG Bo-bo1(1.College of Life Science, Yan’an University, Shanxi Engineering & Technological Research Center for Conversation & Utilization of Regional Biological Resources,Yan’an 716000,China;2.College of Agronomy, Gansu Agricultural University,Lanzhou 730070,China);Construction of a fused gene from three kinds of starch synthases of potato (Solanum tuberosum L.) by using one-step overlap-PCR[J];Journal of Gansu Agricultural University;2010-03
5 WEI Yuan-wen,DENG Zhi-nian,HUANG Cheng-mei,PAN You-qiang,Lü Wei-li,LI Yang-rui(Guangxi Crop Genetic Improvement and Biotechnology Key Lab,Nanning 530007,China);Cloning and sequencing of sugarcane ethylene receptor gene(Sc-ERS)[J];Guangxi Agricultural Sciences;2010-10
6 HUANG Xing,ZENG Xing,LIU Ming,GONG Bo,HU Miao,CAO Zhu-an(Department of Chemical Engineering,Tsinghua University,Beijing 100084,China);Site-directed Mutagenesis of Putative Adenylylation Site of Glutamine Synthetase and Glutamine Production[J];The Chinese Journal of Process Engineering;2008-01
7 DING Ai-jun REN Li-wei FU De-zhiLI Bi-chun(College of Animal Science and Technology,Yangzhou University,Yangzhou 225009,China);Expression of Neuraminidase of H5N1 Subtype Avian Influenza Virus in NIH-3T3 Cells[J];Acta Agriculturae Universitatis Jiangxiensis;2010-04
8 DAI Xin-xin, LI Bian-sheng (College of Light Industry and Food, South China University of Technology, Guangzhou 510640, Guangdong, China);ADVANCES IN SEED ALLERGENS RESEARCH[J];Food Research and Development;2007-07
9 Song Wentao1,2 He Chongbo2 Gao Xianggang2 Liu Xing2,3 Geng Huijun1,2 (1College of Life Sciences,Liaoning Normal University,Dalian 116029;2Liaoning Ocean and Fishery Science Institute, Liaoning Key Laboratory of Marine Fishery Molecular Biology,Dalian 116023);Study on Intron in Fish[J];Biotechnology Bulletin;2009-01
10 ZHANG Qing-hua,HAN Yue-wu,XIE Jun-qiu,et al(Institute of Biochemistry and Molecular Biology,College of Basic Medical Sciences,Lanzhou University,Lanzhou 730000,China);Expression of Antimicrobial Peptide Dermaseptin S4 Gene and Its Mutant in E. coli[J];Chinese Journal of Biologicals;2008-04
【Secondary Citations】
Chinese Journal Full-text Database 10 Hits
1 Ni Bin(College of Life Science Nanjing Agricultural University Nanjing Jiangsu 210095);Current Research Progress of Plant Marker free Transgenic Technology[J];Anhui Agricultural Science Bulletin;2007-16
2 CUI Guang rong (Anhui Technical Teachers College,Fengyang 233100);Methods and Characteristics of Plant Transgene and Gene Silence[J];Journal of Anhui Technical Teachers College;2003-01
3 ZHEN Wei WANG Yangming\ DING Wei\ CHEN Xi\ Hu Yuanlei\ LIN Zhongping (National Laboratory of Protein Engineering and Plant Genetic Engineering,Peking University,Beijing,100871);Cre/loxP-mediated Deletion of Transgene in Transgenic Tobacco[J];Acta Scicentiarum Naturalum Universitis Pekinesis;2001-04
4 Lu Hai; Wu Wei; Zeng Qingyin; Li Yi; Jiang Xiangning; Wang Shasheng. College of Biology, Beijing For. Univ., 100083, P.R.China.\;;The expression analysis of recombinant protein in \%Escherichia coli BL21 (DE3).[J];Journal of Beijing Forestry University;2001-06
5 WANG Wen-qi;GAI Ying;LU Hai;JIANG Xiang-ning.College of Biological Sciences and Biotechnology,Beijing Forestry University,100083,P.R.China.;Recombinase Cre's overexpression in Escherichia coli BL21(DE3) and its one-step purification and activity assay.[J];Journal of Beijing Forestry University;2006-03
6 LI Jian-jun,NI Jing-tao(China Agricultural University,Beijing 100094,China);The Ethical Issues and Moral Responsibilities in the Innovation of Genetically Modified Technology[J];Journal of Baoji University of Arts and Sciences(Social Science Edition);2006-05
7 CHEN Xiao Sui,WANG Bao An,WANG Yan *(Central Laboratory,Navy General Hospital of PLA,Beijing 100037,China);The condition for error-prone PCR induced mutation[J];Academic Journal of Second Military Medical University;2003-03
8 Ge Qingyan Su Qiao An Lijia*Dalian University of Technology, Dalian, 116023* Corresponding author, Bioeng@dlut.edu.cn;The Strategies of Selectable Marker Genes in Transgenic Plants Control[J];Molecular Plant Breeding;2004-05
9 LUO Shi-ping , LENG Xi-gang . Institute of Biomedical Engineering,Chinese Academy of Medical Sciences & Peking Union Medical College,Key Laboratory of Biomedical Materials,Tianjin 300192, China;PCR-based site-directed mutagenesis[J];;2005-03
10 ZHANG Wei.SHEN Bei-fen (Institute of Basic Medical Sciences , Academy of Military Medical Sciences , Beijing 100850, China);A fast and simple method for site-specific mutagenesis[J];Immunological Journal;2003-S1
©2006 Tsinghua Tongfang Knowledge Network Technology Co., Ltd.(Beijing)(TTKN) All rights reserved