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Tineke H. Jones Alain Houde Elyse Poitras Pierre Ward Michael W. Johns 《Food and environmental virology》2009,1(2):57-65
There are increasing concerns of zoonotic transmission of some animal enteric viruses, such as calicivirus, hepatitis E virus,
and rotavirus, which are closely related to human pathogenic strains. Most enteric viruses are detected by molecular techniques
because they cannot be cultured. Surrogates such as F-RNA coliphages are cultivable but few molecular methods exist. Individual
real-time TaqMan RT-PCR assays for the replicase gene of F-RNA coliphage genogroups I and IV were developed and multiplexed
with a real-time TaqMan RT-PCR assay for feline calicivirus as a sample process control for the simultaneous detection and
enumeration of genogroup I and IV F-RNA coliphages. Genogroup IV were successfully detected with the multiplexed assay in
80% of fecal samples that contained F-RNA coliphage levels ≥3.2 log plaque forming units (pfu). F-RNA coliphage were at or
below the limit of detection in most fecal samples when levels were ≤4 log pfu/g. 相似文献
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干扰烟草GA 20-氧化酶siRNA植物表达载体的构建及矮化烟草的产生 总被引:2,自引:0,他引:2
根据烟草(Nicotiana tabacum)GA 20-氧化酶基因序列,设计2对分别含有特定酶切位点的特异引物,以烟草基因组DNA为模板,扩增目的基因(约250 bp)片段.将正、反向目的片段分别插入中间载体的内含子两侧.再经BamH I和Sac I双酶切回收约700 bp的目的片段,插入到双元载体质粒p2355中,成功构建了含GA 20-氧化酶基因片段的反向重复序列植物表达载体p23700,其转录产物能形成发夹RNA(hpRNA),产生小分子干扰RNA,干扰目的基因的表达.将p23700质粒导入根癌农杆菌EHA105中并转化烟草叶片细胞,经选择分化培养,获得表型矮化的转基因烟草.图4参14 相似文献
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利用RNA干涉技术构建水稻DDB1(Damaged DNA binding protein 1)基因不同启动子驱动植物表达载体,DDB1-RNAi和DDB1-glu-RNAi.通过根癌农杆菌介导转入水稻愈伤,经组织培养成功获得转基因植株.半定量RT-PCR分析显示,与野生型植株相比,两组转基因植株幼叶内DDB1的表达量... 相似文献
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以中华绒螯蟹(Eriocheir sinesis)仔蟹为研究对象。采用静水试验法,温度在20℃左右,用氨氮与过滤海水分设5个质量浓度组(2.04、6.35、9.04、11.75和19.13 mg.L-1),同时以过滤海水为对照组(氨氮质量浓度为0.30 mg.L-1),每一个质量浓度组设3个平行样本,对中华绒螯蟹仔蟹进行6 d胁迫试验。每2 d观察仔蟹的蜕壳情况,并分别于实验后的2、4和6 d取仔蟹肝组织样,应用激光共聚焦技术分析测定RNA/DNA荧光相素比,同时用透射电镜观察6 d后,最高浓度组和对照组仔蟹肝细胞结构的变化情况。试验结果表明;对于氨氮的胁迫,仔蟹的变态率随染毒浓度的增高而降低,同时变态蜕壳的时间也会产生延迟现象。氨氮的胁迫会引起仔蟹的肝组织细胞内的DNA和RNA的质量分数逐渐降低,导致RNA/DNA比率的不断下降。在高质量浓度氨氮的胁迫下,使得仔蟹肝细胞线粒体部分解体、胞质空泡化和染色质浓缩、转运泡数量增多,体积增大,细胞出现许多空泡和微绒毛消失等一系列的损伤变化。破坏了仔蟹自身机能正常的代谢和生长水平。 相似文献
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