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《China Biotechnology》 2009-08
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Relative and Absolute Quantification-Expression Analysis of CsSAMDC Gene As a Case

ZHANG Ji-yu WANG Yan-rong NAN Zhi-biao(College of Pastoral Agricultural Science and Technology,Lanzhou University,Lanzhou 730020,China)  
This experiment was based on a full-length SAMDC sequence from the drought stress induced cDNA library of Cleistogenes songorica.Two methods,absolute quantification and relative quantification,were used to analyze the gene expression under different tissue and drought stress treatment by real-time quantitaive PCR experiments and were compared each other,while CsSAMDC gene was used as a case.Leaves and roots tissue were sampled from the plants at 0,4,6,8,and 10 d of drought stress and 1 and 4 d after rewatering.The 2-CT method was used to analyze the relative changes in gene expression from quantitative real-time PCR experiments.A standard curve was generated by the purified plasmid DNA.The primer concentration in the realtime PCR was optimized.All absolute quatification data were normalized by the normalization factor from the most stable house-keeping genes in C.songorica.Relative quantification results using 2-CT method showed that CsSAMDC transcripts increased 5.93±0.71 fold in roots,down regulated 0.62±0.13 fold in leaves,under drought stress.Absolute quantification results showed that CsSAMDC transcripts up-regulated in roots significantly under drought stress as 1ong as 8 to 1 0 days after drought stress,while down-regulated in both roots and leaves after drought stress within 6 days and 1 to 4 days after rewatering,compared with control.CsSAMDC transcipt showed similar tendency of gene expression compared by using relative and absolute quantification methods.
【Fund】: 国家“973”计划(2007CB108904);; 科技部国际科技合作重点项目(2004DFA02000);; 国家科技支撑计划(2008BADB3803);; 甘肃省国际科技合作重点项目计划(4WS064-A71-058)资助项目
【CateGory Index】: Q78
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【Citations】
Chinese Journal Full-text Database 1 Hits
1 ZHANG Chi-Yu**, XU Shun-Gao, HUANG Xin-Xiang(Jiangsu University School of Medical Technology, Zhenjiang 212001, China);A Novel and Convenient Relative Quantitative Method of Fluorescence Real Time RT-PCR Assay Based on Slope of Standard Curve[J];Progress In Biochemistry and Biophysics;2005-09
【Co-citations】
Chinese Journal Full-text Database 10 Hits
1 MA Tao,LEI Yancang,HAO Youhua,WANG Baoju,YANG Dongliang Division of Clinical Immunology,Tongji Hospital,Tongji Medical College,Huazhong University of Science & Technology,Wuhan,430030,China;Cloning and Eukaryotic Expression of APOBEC3F and Its Anti-HBV Effect[J];Journal of Medical Molecular Biology;2007-04
2 LIU Bin DONG Yan SONG Yi ZHENG Xiaofei SUN Zhixian (Institute of Radiation Medicine, Academy of Military Medical Sciences, Beijing 100850, China);Evaluation of the feasibility with Gadd45 as a radiation biodosimeter[J];Journal of Radiation Research and Radiation Processing;2008-03
3 YANG Xiaoyu1,2,YAO Jianfeng1,HUANG Yanfang3,ZHANG Mengzhang4 1.Center of Cell and Developmental Biology,Fujian Medical University,FuZhou 350004,China;2.Fujian Reproductive Medicine Center,Fujian Medical University,FuZhou 350004,China;3.The Affiliatied First Hospital,Fujian Medical University,FuZhou 350005,China;4.Xiamen Entry-Exit Inspection and Quarantine Bureau,Xiamen 361012,China;Detection of Grb10 Gene Expression in Cultured Cells by Real-Time PCR[J];Journal of Fujian Medical University;2008-06
4 DUAN Nai-Rong,CHEN Hong,WANG Qi-Jie Department of Infectious Diseases,First Hospital,Lanzhou University,Lanzhou 730000,China;Correlation between expression of TLR7 mRNA in peripheral blood monouclear cells of chronic hepatitis B patients and IL-12,HBV-DNA load[J];Journal of the Fourth Military Medical University;2009-15
5 SUN Xiao-min,YU Jia-lin,LIN Li-hua,LIN Ya-yin,ZOU Zhi-hui (Department of Neonatology,Children’s Hospital,Chongqing Medical University,Chongqing 400014,China);Effects of ambroxol on the signal molecules and expressions of the related genes of quorum-sensing system in Pseudomonas aeruginosa biofilm[J];Acta Academiae Medicinae Militaris Tertiae;2009-16
6 FANG Zhihui,XU Min,CHENG Kai,ZHAO Yijun(College of Life Science,Huazhong Normal University,Wuhan 430079);Quantification of microcystin synthetase gene mcyH using real-time quantitative RT-PCR with SYBR greenⅠ[J];Journal of Huazhong Normal University(Natural Sciences);2008-03
7 Xu Lijun,Shan Yuxi~*,Chen Dong,Gao Jie,Yang Dongrong,Sun Chuanyang,Cui Yong,Xue Boxin Department of Urology,Affiliated Second Hospital of Soochow University,Suzhou 215004,China;The expression of CD34 and Sca-1 protein in rat penile corpus cavernosum at different-month-old[J];Chinese Journal of Andrology;2009-06
8 NIU Rui1,LIU Bo2,SHAO Ming-ju1,WANG Wei1(1.Department of Respiratory,Second Hospital of Shandong University,Jinan 250033,Shandong,China;2.Department of Respiratory,Weihai Municipal Hospital,Weihai 264200,Shandong,China);mRNA expression of lunx in regional lymph nodes of non-small cell lung cancer and its correlation with prognosis[J];Journal of Shandong University(Health Sciences);2007-09
9 YU Lei1,LIU Jian-ping1,2,ZHUANG Zhi-xiong1,YANG Lin-qing1,ZHANG Ren-li1,YE Xiao-ming1,CHENG Jin-quan1(1.Shenzhen Center for Disease Control and Prevention,Shenzhen 518020;2.School of Public Health,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China);Quantitative Analysis of Real-Time PCR Expression Production by REST and 2~((-ΔΔCT))[J];Journal of Tropical Medicine;2007-10
10 YANG Lin1,ZONG Ming1, LU Tian-bao1,LI Hong-mei3,HAN Jie2,WANG Qiang4,HE Jun-min5,FAN Lie-ying1 (1. Department of Clinical Laboratory;2. Department of Immunology, Dongfang Hospital, Tongji University, Shanghai 200120,China;3. National Engineering Research Center,Shanghai 201203,China;4. Department of Surgery,Changzheng Hospital,Second Military Medical University,Shanghai 200003,China;5. School of Life Science and Technology,Tongji University,Shanghai 200092,China );Expression of FOXP3 and GITR/GITRL mRNA in peripheral blood monocytes of patients with rheumatoid arthritis and colorectal cancer[J];Current Immunology;2008-01
【Secondary Citations】
Chinese Journal Full-text Database 3 Hits
1 Zhang Yan, Shen Qian ** (Department of Central Laboratory, The Affiliated Changhai Hospital, Second Military Medical University, Shanghai 200433,China);Rapid Construction of cRNA Standard Curves in Real-time Reverse Transcription Polymerase Chain Reaction to Measure mRNA Expression[J];Progress In Biochemistry and Biophysics;2003-03
2 ZHANG Chi yu 1), 2) , ZHANG Gao hong 1, 2) , YANG Min 1), BEN Kun long 1) ( 1) Laboratory for Molecular and Cell Immunology, Kunming Institute of Zoology, The Chinese Academy of Sciences, Kunming 650223, China; 2) The Graduate School of Chinese Academy of Sciences, Beijing 100039, China);Elimination of Primer-dimer Effect in SYBR Green Ⅰ Real-time RT-PCR Using 4-step Program[J];Chinese Journal of Biochemistry and Molecular Biology;2004-03
3 XU Shun-gao, HUANG Xin-xiang, ZHOU Li-ping ( Medical Technology School of Jiangsu University, Zhenjiang Jiangsu 212001 , China);A Method of Fluoresce Real time RT-PCR for Investigating the z66 and d or j Antigen Genes Expression of Salmonella Enterica Serovar Typhi[J];Journal of Zhenjiang Medical College;2005-01
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