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| 1 | Gateway重组系统快速构建花生条纹病毒cp基因反向重复序列载体显示文摘Gateway重组系统是Invitrogen公司推出一种新的克隆策略。鉴于一般用于农杆菌转化植物的质粒比较大,采用常规的酶切和连接的克隆策略耗时、费力,Gaytewayw重组系统为大载体的操作提供了一种快速和可靠的手段。该系统利用噬菌体转染细菌后整合细菌DNA的位点特异性重组策略(Landy,1989),重组过程是保守位点特异性DNA链的切割和重接,整个过程中没有DNA的合成。应用Gateway重组系统构建植物反向重复序列表达载体包括两个重组过程:第一,BP重组:目的片段重组至入门载体(Entry clone);第二,LR重组,目的片段自入门载体重组至植物表达载体。本实验室拟利用这一系统构建花生条纹病毒(PStV)红安株系的cp反向重复序列载体,进行烟草转化,期望获得抗性的转基因植株。 | 晏立英 许泽永 陈坤荣 Rob Goldbach Marcel Prins | 2007 | 农业生物技术学报2007,15,2: | 5 |
| 2 | 反向重复RNA介导转基因烟草对花生条纹病毒抗性显示文摘将花生条纹病毒(Peanut stripe virus,PStV)外壳蛋白基因(cp)反向重复序列重组至植物表达载体pK7GW1WG2,通过根癌农杆菌(Agrobacterium tumefaciens)菌株GV3101导入本生烟(Nicotiana benthamiana)。卡那霉素抗性植株5~6叶期摩擦接种PStV,通过症状观察和ELISA检测,T0代转基因烟草植株87%表现免疫,不同抗性系T1代植株的免疫比例为60%~100%,多数抗性系T2代植株免疫比例达到100%。利用PStV cp基因特异探针对转基因植株T1代进行特异小干扰性RNA(siRNA)的Northern blot检测,转基因植株都检测出不同含量的特异siRNA,而非转基因植株未检测到特异siRNA;对病毒接种前的转基因植株进行特异siRNA检测,免疫植株比感病植株特异siRNA含量高;对免疫转基因植株进行不同时期的siRNA检测,植株在接种前、接种15和50d后的叶片中都检测到特异siRNA,同一植株不同时期siRNA含量相对稳定;各个转化系植株体内siRNA含量存在差异。实验成功地利用双链RNA(dsRNA)策略获得对PStV具有较高比例抗性的转基因烟草植株。 | 晏立英 许泽永 陈坤荣 Rob Goldbach Marcel Prins | 2007 | 农业生物技术学报2007,15,4: | 3 |
| 3 | 抗生素作为严重急性营养不良治疗的一部分显示文摘背景:严重急性营养不良每年导致100万儿童死亡。营养治疗中常规加入抗生素可能提高社区中治疗的严重急性营养不良儿童的恢复率,并降低死亡率。 | Trehan I Goldbach HS LaGrone LN 柏振江 汪健 | 2013 | 中华实用儿科临床杂志2013,28,11: | 3 |
| 4 | 侵染花生的黄瓜花叶病毒CA株核酸全序列分析显示文摘对侵染花生的黄瓜花叶病毒CA(CMVCA)株系进行克隆和序列分析。CMVCARNA1全长3356个核苷酸(nt),编码分子量为111kDa的1a蛋白;RNA2全长3045nt,编码分子量为96.7kDa的2a蛋白和13.1kDa的2b蛋白;RNA3全长2219nt,编码分子量为30.5kDa的3a蛋白和分子量为24kDa的外壳蛋白(CP)。序列同源性比较表明,CMVCARNA1、2、3与CMV亚组IACMVFny、亚组IBCMVSD、亚组IICMVQ株系序列同源性,RNA1分别为91.3%、91.1%和76.5%,RNA2分别为92.1%、90%和71.2%,RNA3分别为96.1%、92.6%和74.5%;与同属花生矮化病毒(Peanutstuntvirus,PSV)RNA1、2、3序列同源性分别为67.1%、58.2%和55.7%。上述研究未发现CMVCA基因组与PSVRNA链的重组,CMVCA对花生的侵染应是该株系适应于花生的遗传变异、长期进化的结果;对RNA35′NTR结构分析以及RNA35′NTR和CP系统进化树分析表明,CMVCA属CMVIB亚组。 | 晏立英 许泽永 陈坤荣 Rob Goldbach Marcel Prins | 2005 | 中国病毒学2005,20,3: | 3 |
| 5 | Pulse Electrodeposition and Electrochemical Quartz Crystal Microbalance Techniques for High Perpendicular Magnetic Anisotropy Cobalt Nanowire Arrays显示文摘 | Ursache A Goldbach J T Tuominen M T | 2005 | J Appl Phys2005,97,10: | 1 |
| 6 | Boron is required for the ferricyanide-induced proton release by auxins in suspension-cultured cells of Daucus carota and Lycopersicon esculentum显示文摘 | Goldbach H E Hartmann D Rotzer T | 1990 | Physiol Plant1990,80,: | 1 |
| 7 | Defect sealing in Pd membranes via point plating 显示文摘 | ZENG G F GOLDBACH A XU H Y | 2009 | Journal of Membrane Science2009,328,1: | 1 |
| 8 | Boron-and-salt interactions in wheat areaffected by boron supply显示文摘 | Wimmer M A Goldbach H E | 2012 | Journal of Plant Nutrition and Soil Science、2012,175,2: | 1 |
| 9 | A critical review of current hypothesis concerning the role of boron in higher plant:suggestion for further research and methodological显示文摘 | Goldbach H E | 1997 | J Trace Microp Tech1997,15,1: | 1 |
| 10 | Sterile filtration of liposomes: retention of encapsulated carboxyfluorescein 显示文摘 | Goldbach P Brochart H Wehrle P | 1995 | Intenlational Journal of Pharmaceutics1995,117,: | 1 |
| 11 | Effect of choice of measles mumps - rubella vaccine on immediate vaccination pain in infants显示文摘 | Ipp M Cohen E Goldbach M Macarthur C | 2004 | Arch Pediatr Adolesc Med2004,158,6: | 1 |
| 12 | Tomato spotted wilt virus infection improves host suitability for its vector Frankliniella occidentalis 显示文摘 | Maris PC Joosten NN Goldbach RW | 2004 | Phytopathology2004,94,7: | 1 |
| 13 | Assessing the expression of chicken anemia virus proteins in plants 显示文摘 | Lacorte C Lohuis H Goldbach R | 2007 | Vir Res2007,129,2: | 1 |
| 14 | High frequency induction of RNA-mediated resistance against Cucumber mosaic virus using inverted repeat constructs 显示文摘 | Chen Y K Lohuis D Goldbach R | 2004 | Molecular Breeding2004,14,: | 1 |
| 15 | Genome similarities between plant and an- imal RNA viruses显示文摘 | Goldbach R | 1987 | Microbiol Sci1987,4,: | 1 |
| 16 | A critical review on current hypotheses concerning the role of boron in higher plants: suggestions for further research and methodological requirements显示文摘 | Goldbach H E | 1997 | Joumal of trace and microprobe techniques1997,15,1: | 1 |
| 17 | RNA-mediated virus resistance in transgenic plants显示文摘 | PRINS M GOLDBACH R | 1996 | Archives of Virology1996,141,12: | 1 |
| 18 | Evolution of plus-strand RNA viruses显示文摘 | Goldbach R Wellink J | 1988 | Intervirology1988,29,: | 1 |
| 19 | RNA-mediated virus resistance in transgenic plants显示文摘 | GOLDBACH R | 1996 | Arch Virol1996,141,: | 1 |
| 20 | A critical review on current hypotheses concerning the role of boron in higher plants:suggestions for further research and methodological requirements显示文摘 | Goldbach H E | 1997 | J T race and Microprobe Techniques1997,15,1: | 1 |