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《Carcinogenesis,Teratogenesis and Mutagenesis》 2005-02
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Influence of Plasmid Preparation and Temperature on Nested Deletions

WANG Zi-liang 1 ,XU Li-yan 2 ,LI En-min 1, (1.Department of Biochemistry and Molecule Biology,College of Medicine Science,Shantou University,Shantou Guangdong515041,China;2.Department of Pathology,College of Medicine Science,Shantou University,Shantou Guangdong515041,China)  
BACKGROUNDAIM: To explore the influencing factors of nested deletions and find some methods to resolve interrelated problems. MATERIAL AND METHODS: Alkaline lysis,purification by polyethylene glycol(PEG)and QIAprep Spin Miniprep Kit were used to prepare the plasmid pGLB-G6carrying the5'flanking region of NGAL gene to study the nonspecific degradation of Exonuclease(ExoⅢ),the best method of preparing the plasmid,not susceptible to ExoⅢ,would be screened.ExoⅢdeletion reaction was generated at22℃and37℃respectively and the agarose gel was run to determine the extent of digestion.PCR amplification and restriction endonuclease digestion was used to identify deleted DNA fragments. RESULTS: The plasmid extracted by QIAprep Spin Miniprep Kit is insusceptible to ExoⅢand suitable for deletion reaction.The agarose gel electrophoresis showed that DNA fragments had not obviously shortened at22℃,but at37℃.Restriction endonuclease digestion is needed for further the identification of deleted DNA fragments. CONCLUSION: The quality of plasmid DNA is a crucial factor to generate specific deletion reactions of Exo III,the plamid extracted by QIAprep Spin Miniprep Kit is satisfactory;setting a control of37℃may ensure deletion reactions have happened and by PCR amplification and restriction endonuclease digestion,real deleted DNA can be attained.
【Fund】: 国家自然科学基金面上项目(No.39900069 No.30170428 No.30370641);; 广东省自然科学基金重点项目(No.37788) ;; 广东省自然科学基金面上项目(990799 010431)
【CateGory Index】: Q784
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【Citations】
Chinese Journal Full-text Database 3 Hits
1 XU La - yan, LI En - min, CAI Wei - jia , et al (Department of Oncologic Pathology, Medical College of Shantou University, Shantou 515031,China);Constructing and Identifying the Firefly Luciferase Report Gene Vectors For 5' Flanking Regulated Elements of NGAL Gene[J];Carcinogenesis,Teratogenesis and Mutagenesis;2004-01
2 XU Li Yan 1)** , LI En Min 2) , XIONG Hua Qi 1) , CAI Wei Jia 1) , SHEN Zhong Ying 1) ( 1) Institute of Oncologic Pathology, Medical College of Shantou University, Shantou 515031, China; 2) Department of Bioche;Study of Neutrophil Gelatinase-assiciated Lipocalin(NGAL) Gene Overexpression in the Progress of Malignant Transformation of Human Immortalized Esophageal Epithelial Cell[J];Progress In Biochemistry and Biophysics;2001-06
3 LI En-Min 1, 2*, XU Li-Yan 2, CAI Wei-Jia 2, XIONG Hua-Qi 2, SHEN Zhong-Ying 2, ZENG Yi 3 ( 1Department of Biochemistry and Molecular Biology, Shantou University Medical College, Shantou 515031, China; 2Institute of Oncologic Pathology, Shantou University Medical College, Shantou 515031, China; 3Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052, China );Functions of Neutrophil Gelatinase-associated Lipocalin in the Esophageal Carcinoma Cell Line SHEEC[J];Acta Biochimica Et Biophysica Sinica;2003-03
【Co-citations】
Chinese Journal Full-text Database 10 Hits
1 NIU Yong_dong 1, XU Li_yan 2, LI En_min 1(1. Department of Biochemistry & Molecular Biology; 2. Insititue of Oncologic Pathology,Medical College of Shantou University, Shantou 515031 , China);CONSTRUCTION AND IDENTIFICATION OF THE BAIT PLASMIDS pHis-3NF AND pLacZ-3NF FOR YEAST ONE-HYBRID SYSTEM[J];Carcinogenesis,Teratogenesis and Mutagenesis;2003-01
2 XU La - yan, LI En - min, CAI Wei - jia , et al (Department of Oncologic Pathology, Medical College of Shantou University, Shantou 515031,China);Constructing and Identifying the Firefly Luciferase Report Gene Vectors For 5' Flanking Regulated Elements of NGAL Gene[J];Carcinogenesis,Teratogenesis and Mutagenesis;2004-01
3 WANG Zhao_yang 1 ,XU Li_yan 1,ˇ ,RONG Ju 2 ,et al (1.Insititute of Oncologic Pathology,Medical college of Shantou University,Shantou515031,China;2.Department of Biochemistry&molecularBiology,Medical CollegeofShantouUniversity,Shantou515031,China);Purification by Ni~(2+)-Metal Chelate Affinity Chromatography and Identification of NGAL Protein[J];Carcinogenesis,Teratogenesis and Mutagenesis;2004-02
4 XIE Yong-fang, SHU Kun-xian, LIANG Yi-long, et al(Biology College, Chongqing University ofPosts and Communications, Chongqing 400065, China);Cell Proliferation and Apoptosis in the Developing Intestine of Mouse[J];Carcinogenesis,Teratogenesis and Mutagenesis;2005-01
5 KANG Zhen-qiao1, JI Yan-xia2 , YANG Zhi-yong1,SU Guang1,CUI Yan-zhi3 (1. Handan the Second Hospital,056001, Hebei, China; 2. Handan Central Hospital, Handan 056001, Hebei, China;3. Shijiazhuang the First Hospital, Shijiazhuang 050011, Hebei, China);The Expression of Vascular Endothelial Growth Factor in Breast Cancer Cell Lines[J];Carcinogenesis,Teratogenesis & Mutagenesis;2006-03
6 AN Gang, HUANG Tian-hua, XIE Qing-dong, WANG De-gang.(Research Center for Reproductive Medicine, Shantou University Medical College, Shantou 515041,China);Effect of pCXN2-mIZUMO on the Fertility of C57BL/6 Mice[J];Carcinogenesis,Teratogenesis & Mutagenesis;2007-04
7 ZHU Bin,YANG Luo-yan,ZHAO Xiao-kun(Department of Urology,Xiangya Second Hospital of Central South University,Changsha 410011,China);Relationship Between Expressions of MT-3 and MDR-1 in Bladder Cancer[J];Carcinogenesis,Teratogenesis & Mutagenesis;2007-05
8 WU Bing-li1,3,XU Li-yan2,YING Xiao-min3,NIU Yong-dong1,LI Wu-ju3,,LI En-min1,(1.Biochemistry and Molecular Biology Department,Medical College of Shantou University,Shantou 515041;2.Department of Tumor Pathology,Medical College of Shantou University,Shantou 515041;3.Institute of Basic Medical Sciences,Academy of Military Medical Sciences,Beijing 100850,China);Silicon Cloning for the Unknown Functional Genes Related to Esophageal Carcinoma and ncRNA Finding[J];Carcinogenesis,Teratogenesis & Mutagenesis;2008-02
9 Bi Yuping,Liao Junjie,Shan Lei,Wang Xingjun & Xu Pingli(Biotechnology reseacher center,Shandong Academy of Agricultural Sciences,Jinan 250100);Study on Transgenic Tomato Plants Expressing Resistance to Cucumber Mosaic Virus and Tobacco Mosaic Virus[J];Shandong Agricultural Sciences;1997-06
10 Shan Lei,Bi Yuping,Wang Xingjun,Xu Pingli & Li Guangcun (Biotechnology Research Center,Shangdong Academy of Agricultural Sciences,Jinan 250100);Genetic Analysis on the Progeny of the Transgenic Tomato with Double-resistance to TMV and CMV[J];SHANDONG AGRICULTURAL SCIENCES;1998-04
China Proceedings of conference Full-text Database 10 Hits
1 ;Screening and identification of genes transactivated by XTP3 protein by microarray assay[A];[C];2004
2 WANG Xiao-qian TANG Nan-hong LI Xiu-jin,Hepato-Biliary Surgery Institute of Fujian Province,Union Hospital Affiliated to Fujian Medical University,Fuzhou 350001,China;Construction of hepatocytes with specific high-expression of human Glutamine Synthetase (hGS)[A];[C];2007
3 ;子房注射法获得转基因甜瓜植株[A];[C];2006
4 SHANG Qing-long, et al (Microbiol Teach & REs . Sect. Harbin med. Univ. Harbin 150086);Expression, purification, and immunization of HPV16 L1 protein in E .coli[A];[C];2003
5 Zhenwen Chen, Taiyun Zhao, Hejuan Sun, Lujing, Wangying, Mengxia, Zhenghe Chao, Wangju (Veterinary Medical College of China Agriculture University, Beijing 100094; Department of Laboratory Animal Sciences Capital University of Medical Sciences Laboratory Animal Center of the Academy MilitarMedicalSciences);Comparative analyzing the population genetic of Mongolia gerbil with microsatellite DNA and biochemistry marker[A];[C];2006
6 WANG Ge, WANG Yan, HE Ling-bing,QIAO Gang,ZHU Yi-song, WANG Jun-yan (ShandongYounglong Biotech. Co, Ltd.& Interlong Biotech.,Jinan250101,China);Expression and secretion of recombinant human nerve growth factor in yeast[A];[C];2006
7 REN Hua, GU Jian-hua, SUN Zhu-yun, SHEN Wan-yun, WU Ying, ZHOU Zhong-liang School of Life Science, East China Normal University, Shanghai, 200062;A Reverse Transcription-Polymerase Chain Reaction for Zebrafish(Danio rerio) vitellogenin mRNA[A];[C];2005
8 MA Jun,CHEN Yun-shuang,WANG Ai-shan,Ll Zhong-kui Shanghai Zoo, Shanghai, 200335, China;Identification of the sex of the rare bird of Oriental white stork[A];[C];2005
9 MA Xun LU Cheng-ping(Key Laboratory of Animal Disease Diagnostic and Immunology,Ministry of Agriculture,Nanjing Agricultural University,Nanjing,Jiangsu 210095,China);Analysis on the Complete Genome of Swine Hepatitis E Virus Isolated from Xinjiang[A];[C];2005
10 ;EXPRESSION AND IDENTIFICATION OF IBDV VPS STRUCTURAL PROTEIN IN E. Coli[A];[C];2005
【Secondary Citations】
Chinese Journal Full-text Database 6 Hits
1 Li En Min1, Xu Li Yan2, Xiong Hua Qi2, Cai Wei Jia2, Wu Bing Li1,Zhang Can1, Zhang Yong Fa1, Lin Yan1, Shen Zhong Ying21.Department of Biochemistry andMolecular Biology, 2.Institute of Oncologic Pathology, ShantouUniversity Medical College,Shantou,Guangdong,515031,P.R.China;Cloning and Identification of 5-′Untranslated Region (UTR)and 3′-Untranslated Region of Neutrophil Gelatinase-Associated Lipocalin (NGAL) Gene from Esophageal Carcinoma Cell Line SHEEC[J];Chinese Journal of Cancer;2003-02
2 Shen Zhongying 1 Cai Weijia 1 Shen Jian 1 Xu Jinjie 1 Cen Shan 2 Ten Zhipin 2 Hu Zhi 1 Zeng Yi 2;HUMAN PAPILLOMA VIRUS 18E6E7 IN SYNERGY WITH TPA INDUCED MALIGNANT TRANSFORMATION OF HUMAN EMBRYONIC ESOPHAGEAL EPITHELIAL CELLS[J];CHINESE JOURNAL OF VIROLOGY;1999-01
3 LI En min,XU Li yan,WU Bing li,et al.Department of Biochemistry and Molecular Biology,Medical College of Shantou University,Shantou 515031,China;Cloning of NGAL Gene 5′-nontranscribed Region and Analysis of its Cis-acting Elements in Esophageal Carcinoma Cell SHEEC[J];Journal of Qilu Oncology;2003-03
4 XU Li Yan 1)** , LI En Min 2) , XIONG Hua Qi 1) , CAI Wei Jia 1) , SHEN Zhong Ying 1) ( 1) Institute of Oncologic Pathology, Medical College of Shantou University, Shantou 515031, China; 2) Department of Bioche;Study of Neutrophil Gelatinase-assiciated Lipocalin(NGAL) Gene Overexpression in the Progress of Malignant Transformation of Human Immortalized Esophageal Epithelial Cell[J];Progress In Biochemistry and Biophysics;2001-06
5 LI En-Min 1, 2*, XU Li-Yan 2, CAI Wei-Jia 2, XIONG Hua-Qi 2, SHEN Zhong-Ying 2, ZENG Yi 3 ( 1Department of Biochemistry and Molecular Biology, Shantou University Medical College, Shantou 515031, China; 2Institute of Oncologic Pathology, Shantou University Medical College, Shantou 515031, China; 3Institute of Virology, Chinese Academy of Preventive Medicine, Beijing 100052, China );Functions of Neutrophil Gelatinase-associated Lipocalin in the Esophageal Carcinoma Cell Line SHEEC[J];Acta Biochimica Et Biophysica Sinica;2003-03
6 SHGN Zhongying, CEN Shan, ZENG Yi, et al.Medical College of Shantou University, Shantou 515031;Immortalization of human fetal esophageal epithelial cells induced by E 6 and E 7 genes of human papilloma virus 18[J];CHINESE JOURNAL OF EXPERIMENTAL AND CLINICAL VIROLOGY;1999-02
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