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The intracellular mechanism of alpha-fetoprotein promoting the proliferation of NIH 3T3 cells 被引量:27
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作者 MENG SEN LI, PING FENG LI, FBI YI YANG, SHI PENG HE, Guo GUANG DU, GANG LI1 Department of Biochemistry and Molecular Biology, 2 Department of Biophysics, Health Science Center, Peking University, Beijing 100083, China 《Cell Research》 SCIE CAS CSCD 2002年第2期151-156,共6页
AIM: The existence and properties of alpha-fetoprotein (AFP) receptor on the surface of NIH 3T3 cells and the effects of AFP on cellular signal transduction pathway were investigated. METHODS: The effect of AFP on the... AIM: The existence and properties of alpha-fetoprotein (AFP) receptor on the surface of NIH 3T3 cells and the effects of AFP on cellular signal transduction pathway were investigated. METHODS: The effect of AFP on the proliferation of NIH 3T3 cells was measured by incorporation of 3H-TdR. Receptor-binding assay of 125I-AFP was performed to detect the properties of AFP receptor in NIH 3T3 cells. The influences of AFP on the [cAMP]i and the activities of protein kinase A (PKA) were determined. Western blot was used to detect the change of K-ras P21 protein expression. RESULTS: The proliferation of NIH 3T3 cells treated with 0-80 mg/L of AFP was significantly enhanced. The Scatchard analysis indicated that there were two classes of binding sites with KD of 2.722 x 10(-9)M (Bmax=12810 sites per cell) and 8.931 x 10(-8)M (Bmax=119700 sites per cell) respectively. In the presence of AFP (20 mg/L), the content of cAMP and activities of PKA were significantly elevated . The level of K-ras P21 protein was upregulated by AFP at the concentration of 20 mg/L. The monoclonal antibody against AFP could reverse the effects of AFP on the cAMP content, PKA activity and the expression of K-ras p21 gene. CONCLUSION: The effect of AFP on the cell proliferation was achieved by binding its receptor to trigger the signal transduction pathway of cAMP-PKA and alter the expression of K- ras p21 gene. 展开更多
关键词 3T3 Cells Animals Cell Division Cyclic AMP Cyclic AMP-Dependent Protein Kinases Dose-Response Relationship Drug Humans Mice Receptors Peptide Research Support Non-U.S. Gov't Signal Transduction time Factors ALPHA-FETOPROTEINS
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初级汉语水平韩国留学生的时间词使用偏误分析 被引量:11
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作者 刘慧清 《暨南大学华文学院学报》 2005年第3期19-27,共9页
本文从教学实践出发,对初级汉语水平韩国留学生时间词的误用现象作了一些梳理,总结了一些偏误类型,包括时间词与其他词语顺序错误(时间词作状语位置不当,时间词作补语、定语位置不当,时间词排列顺序错误),时点词、时段词使用混淆,时间... 本文从教学实践出发,对初级汉语水平韩国留学生时间词的误用现象作了一些梳理,总结了一些偏误类型,包括时间词与其他词语顺序错误(时间词作状语位置不当,时间词作补语、定语位置不当,时间词排列顺序错误),时点词、时段词使用混淆,时间词与其他时态成分表时不一致。文章还分析了偏误的原因,给出了修改方法及教学上的应对策略。 展开更多
关键词 时间词 时点词 时段词 后置时间词
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刺五加鲨烯合酶基因家族两成员的表达及其与皂苷含量的关系 被引量:15
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作者 邢朝斌 龙月红 +3 位作者 李非非 周秘 修乐山 劳凤云 《西南农业学报》 CSCD 北大核心 2014年第3期1252-1255,共4页
为了探明刺五加鲨烯合酶(SS)基因家族2成员对皂苷含量的作用机制,以actin为内参基因,利用real time PCR技术,分析刺五加不同生长发育时期、不同器官及茉莉酸甲酯(MeJA)处理对SS1、SS2基因表达及皂苷含量的影响。结果表明:刺五加SS基因... 为了探明刺五加鲨烯合酶(SS)基因家族2成员对皂苷含量的作用机制,以actin为内参基因,利用real time PCR技术,分析刺五加不同生长发育时期、不同器官及茉莉酸甲酯(MeJA)处理对SS1、SS2基因表达及皂苷含量的影响。结果表明:刺五加SS基因家族的SS1和SS2在整个生长期和各器官中均有表达,且表达量差异显著(P<0.05),其中,在萌芽期两者的表达量差异最大。SS1在叶片、叶柄和根器官中的表达量显著高于SS2的表达量(P<0.05)。MeJA处理可显著提高SS1和SS2基因的表达量。刺五加SS1的表达量与皂苷含量间的相关系数低,未达显著水平。SS2的表达量与皂苷含量呈显著的正相关关系(P<0.01)。SS2表达量的高低决定了皂苷含量的高低,是刺五加中三萜皂苷生物合成中的关键酶。 展开更多
关键词 刺五加 表达 鲨烯合酶基因 REAL time PCR 皂苷
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胃癌组织miRNA差异表达的初步分析 被引量:13
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作者 阳圣 张雯 +3 位作者 杨燕青 张小燕 郜恒骏 张庆华 《上海交通大学学报(医学版)》 CAS CSCD 北大核心 2010年第11期1317-1323,共7页
目的分析胃癌组织特异性微小RNA(miRNA)表达谱,为深入研究miRNA与胃癌发生和发展的关系以及寻找新的胃癌分子标志物奠定基础。方法利用miRNA芯片就21对胃癌和癌旁配对正常组织的miRNA表达谱进行检测和初步生物信息学分析,Real-TimePCR验... 目的分析胃癌组织特异性微小RNA(miRNA)表达谱,为深入研究miRNA与胃癌发生和发展的关系以及寻找新的胃癌分子标志物奠定基础。方法利用miRNA芯片就21对胃癌和癌旁配对正常组织的miRNA表达谱进行检测和初步生物信息学分析,Real-TimePCR验证miRNA芯片检测结果。结果 miRNA芯片检测发现,在胃癌及其癌旁配对正常组织中共有88个差异表达miRNA,其中上调最明显的是miR-21、miR-196b、miR-301a、miR-431*、miR-550*、miR-18a和miR-135b;下调最明显的是miR-139-3p、miR-628-3p、miR-596、miR-99b*和miR-638。Real-TimePCR对其中2个上调(miR-181、miR-19b)和2个下调(miR-134、miR-31)miRNA的验证结果与芯片检测所示具有较好的一致性。结论胃癌组织具有特异性的miRNA表达谱,这些差异表达的miRNA有可能成为新的胃癌分子标志物。 展开更多
关键词 微小RNA 表达谱 胃癌 基因芯片 REAL-time PCR
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蝴蝶兰抗坏血酸过氧化物酶基因克隆及其表达研究 被引量:13
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作者 许传俊 孙叙卓 +4 位作者 李玲 茹志伟 曾碧玉 刘育梅 黄珺梅 《园艺学报》 CAS CSCD 北大核心 2012年第4期769-776,共8页
从蝴蝶兰(Phalaenopsis)中克隆获得了抗坏血酸过氧化物酶基因(APX)同源序列,命名为PhAPX(GenBank登录号为:FJ161977)。PhAPX cDNA全长为1320bp,完整的编码框为747bp,编码249个氨基酸。生物信息学分析结果表明,PhAPX属于过氧化物酶家族Cl... 从蝴蝶兰(Phalaenopsis)中克隆获得了抗坏血酸过氧化物酶基因(APX)同源序列,命名为PhAPX(GenBank登录号为:FJ161977)。PhAPX cDNA全长为1320bp,完整的编码框为747bp,编码249个氨基酸。生物信息学分析结果表明,PhAPX属于过氧化物酶家族ClassⅠ的成员,PhAPX蛋白可能是胞质型APX,与其他植物的APX相似性较高。real-time PCR分析表明PhAPX是一个广谱表达的基因,在蝴蝶兰根、茎、叶、花等各个部位都有表达。机械伤害和盐处理都可以诱导PhAPX表达上调,表明PhAPX在胁迫防御中起作用。 展开更多
关键词 蝴蝶兰 APX 克隆 表达 REAL-time PCR
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左归丸对去卵巢大鼠骨髓间充质干细胞基因表达谱时间序列的影响 被引量:11
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作者 赵旭 郝传铮 +1 位作者 杨仁旭 徐俊伟 《中医杂志》 CSCD 北大核心 2015年第16期1414-1419,共6页
目的通过分析骨质疏松模型大鼠骨髓间充质干细胞(BMSCs)基因表达谱时间序列变化趋势,在基因组学层面探索左归丸防治绝经后骨质疏松症的分子机制。方法 72只大鼠随机分为假手术组、模型组、戊酸雌二醇组和左归丸组,每组18只。除假手术组... 目的通过分析骨质疏松模型大鼠骨髓间充质干细胞(BMSCs)基因表达谱时间序列变化趋势,在基因组学层面探索左归丸防治绝经后骨质疏松症的分子机制。方法 72只大鼠随机分为假手术组、模型组、戊酸雌二醇组和左归丸组,每组18只。除假手术组外其余各组大鼠摘除双侧卵巢建立绝经后骨质疏松症模型。手术2周后,戊酸雌二醇组和左归丸组分别给予戊酸雌二醇片溶液(浓度为0.01 mg/ml)和左归丸溶液(浓度为0.5 g/ml)灌胃,假手术组和模型组用等量蒸馏水灌胃,每次1 ml/100 g,每日1次,灌胃至手术后第4、8和12周末(每个时间点6只大鼠)。12周末时检测各组大鼠股骨远端骨密度、胫骨近端骨密、腰椎骨密度。无菌分离各组大鼠的股骨和胫骨,培养BMSCs,表达谱芯片检测各组BMSCs F2代细胞基因表达情况,采用SPSS18.0软件筛选差异表达基因(DEGs),并采用STEM法进行时间序列分析,通过DAVID进行功能与通路富集。结果模型组大鼠股骨远端骨密度、胫骨近端骨密、腰椎骨密度均显著低于假手术组(P<0.01);左归丸组和戊酸雌二醇组大鼠3个骨骼位点的骨密度均高于模型组(P<0.05或P<0.01)。左归丸组-模型组对比组中时间序列显著趋势DEGs多达56个。其中第4周末显著下调基因Ppig、Rb1cc1、Il6和Rock1富集到细胞增殖、自噬凋亡、信号通路和细胞分化等。结论左归丸可能通过影响细胞增殖、自噬凋亡、信号通路和细胞分化等多途径,广泛调控BMSCs基因表达,最终减少骨量丢失。 展开更多
关键词 绝经后骨质疏松症 骨髓间充质干细胞 左归丸 基因表达谱 时间序列 富集分析
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谈“S+T+没(有)+V+(O)+了”句式 被引量:8
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作者 孟艳丽 《汉语学习》 CSSCI 北大核心 2006年第1期38-43,共6页
本文主要讨论三个问题:第一,“S+T+没(有)+V+(O)+了”句式中时量成分为什么一定要放在动词之前?第二,这种句式为什么没有相应的肯定句?第三,为什么在这种句式中“没(有)”和“了”可以共现?文章认为,否定有两种:一种是对量的否定,一种... 本文主要讨论三个问题:第一,“S+T+没(有)+V+(O)+了”句式中时量成分为什么一定要放在动词之前?第二,这种句式为什么没有相应的肯定句?第三,为什么在这种句式中“没(有)”和“了”可以共现?文章认为,否定有两种:一种是对量的否定,一种是对质的否定。时间词语前置,主要是为了区别这两种不同性质的否定。正因为时量词语的前置,使得句子的语序变成了有标记的、句子的否定也变成了有标记的否定,因此没有相应的肯定形式;也正因为时量词语的前置,使得“没(有)”与“了”变成了非直接成分,因此可以在一个句子中共现。 展开更多
关键词 时量词语 量的否定 质的否定 主观长时量
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施氮量和采收时期对油蔬两用型油菜菜薹维生素合成的影响 被引量:9
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作者 林宝刚 怀燕 +5 位作者 李志平 柳寒 任韵 张冬青 余华胜 华水金 《核农学报》 CAS CSCD 北大核心 2020年第12期2813-2822,共10页
为明确施氮量和采收时期对油蔬两用型油菜薹维生素含量的影响,以富硒1号为材料,采用裂区试验设计,设置3个施氮量(120、180和240 kg·hm^-2)和4个采收时期(以株高20、30、40和50 cm表示),分析了油菜薹维生素C、维生素E、维生素B1和... 为明确施氮量和采收时期对油蔬两用型油菜薹维生素含量的影响,以富硒1号为材料,采用裂区试验设计,设置3个施氮量(120、180和240 kg·hm^-2)和4个采收时期(以株高20、30、40和50 cm表示),分析了油菜薹维生素C、维生素E、维生素B1和维生素B6含量的变化;比较了半乳糖内酯脱氢酶(GalLDH)、抗坏血酸氧化酶(AAO)、抗坏血酸过氧化物酶(APX)、脱氢抗坏血酸还原酶(DHAR)和单脱氢抗坏血酸还原酶(MDHAR)活性,并分析了维生素家族基因在菜薹中的表达量。结果表明,随着施氮量的增加,油菜薹还原型维生素C(AsA)含量降低;氧化型维生素C(DHA)含量在株高20 cm采收时增加,但在30 cm及以上株高采收时则降低;维生素E和维生素B6含量先增加后降低;维生素B1含量增加;维生素C合成关键酶GalLDH活性先略增加后降低;AAO和DHAR活性增加,APX和MDHAR活性则整体表现为先增加后降低。且施氮量增加后,维生素家族各基因在株高30和40 cm采收的菜薹中主要表现为正向调控,而在株高20和50 cm采收的菜薹中主要表现为负向调控。就采收时期而言,菜薹中的维生素含量在株高30~40 cm时高于其他时期;在同一施氮量下,随着菜薹采收株高的增加,维生素家族基因的表达量主要呈上调模式。综合分析可知,GalLDH活性的降低以及AAO/DHAR和APX/MDHAR代谢平衡是增施氮肥导致菜薹维生素C含量降低的重要原因。本研究结果为油蔬两用型油菜薹优质栽培技术提供了理论依据。 展开更多
关键词 氮肥 采收时期 油菜薹 维生素 基因表达
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THE RECURSIVE SOLUTION OF QUEUE LENGTH FOR Geo/G/1 QUEUE WITH N-POLICY 被引量:8
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作者 Chuanyi LUO Yinghui TANG +1 位作者 Wei LI Kaili XIANG 《Journal of Systems Science & Complexity》 SCIE EI CSCD 2012年第2期293-302,共10页
This paper considers a discrete-time queue with N-policy and LAS-DA(late arrival system with delayed access) discipline.By using renewal process theory and probability decomposition techniques,the authors derive the r... This paper considers a discrete-time queue with N-policy and LAS-DA(late arrival system with delayed access) discipline.By using renewal process theory and probability decomposition techniques,the authors derive the recursive expressions of the queue-length distributions at epochs n^-,n^+,and n.Furthermore,the authors obtain the stochastic decomposition of the queue length and the relations between the equilibrium distributions of the queue length at different epochs(n^-,n^+,n and departure epoch D_n). 展开更多
关键词 Discrete-time queue N-POLICY recursive expression stochastic decomposition.
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Real-Time PCR Technique and Its Application in Quantification of Plant Nucleic Acid Molecules 被引量:8
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作者 刘进元 《Acta Botanica Sinica》 CSCD 2003年第6期631-637,共7页
Real-time PCR is a closed DNA amplification system that skillfully integrates biochemical, photoelectric and computer techniques. Fluorescence data acquired once per cycle provides rapid absolute quantification of ini... Real-time PCR is a closed DNA amplification system that skillfully integrates biochemical, photoelectric and computer techniques. Fluorescence data acquired once per cycle provides rapid absolute quantification of initial template copy numbers as PCR products are generated. This technique significantly simplifies and accelerates the process of producing reproducible quantification of nucleic acid molecules. It not only is a sensitive, accurate and rapid quantitative method, but it also provides an easier way to calculate the absolute starting copy number of nucleic acid molecules to be tested. Together with molecular bio-techniques, like microarray, real-time PCR will play a very important role in many aspects of molecular life science such as functional gene analysis and disease molecular diagnostics. This review introduces the detailed principles and application of the real-time PCR technique, describes a recently developed system for exact quantification of AUX/IAA genes In Arabidopsis, and discusses the problems with the real-time PCR process. 展开更多
关键词 real-time PCR technique quantification of plant nucleic acid molecules gene expression molecular medicine
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MEF2a基因在肌肉组织中的表达研究 被引量:6
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作者 高智晟 杨龙 +2 位作者 张冬杰 刘娣 刘玉芬 《中国农学通报》 CSCD 北大核心 2009年第4期21-23,共3页
分别取90日龄、180日龄和270日龄大白猪心肌、背肌和腿肌,提取总RNA,设计并合成猪MEF2a引物,以β肌动蛋白作为内参,利用Real-time PCR方法检测不同日龄大白猪肌肉组织中MEF2a基因mRNA的表达。结果表明,不同日龄大白猪的心肌和背肌中,ME... 分别取90日龄、180日龄和270日龄大白猪心肌、背肌和腿肌,提取总RNA,设计并合成猪MEF2a引物,以β肌动蛋白作为内参,利用Real-time PCR方法检测不同日龄大白猪肌肉组织中MEF2a基因mRNA的表达。结果表明,不同日龄大白猪的心肌和背肌中,MEF2a基因mRNA的表达水平无显著差异,在腿肌中有显著差异(P<0.05)。从90日龄到270日龄,MEF2a基因在心肌、背肌和腿肌的表达水平均差异显著(P<0.05),腿肌最高,其次是背肌,心肌最低。据此推测,MEF2a基因在猪成体后的肌肉组织中,主要是促进Ⅰ型肌纤维的生成,而不再是介导细胞分化。 展开更多
关键词 MEF2a基因 肌肉发生 REAL-time PCR
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小麦过氧化物还原酶基因TaPrx的克隆与功能初步分析 被引量:6
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作者 张海燕 李国田 +6 位作者 王晓杰 段迎辉 徐亮胜 郭军 马金彪 黄丽丽 康振生 《中国农业科学》 CAS CSCD 北大核心 2009年第4期1222-1229,共8页
【目的】克隆TaPrx基因并对其在小麦与条锈菌互作中的功能进行初步分析。【方法】利用PCR方法结合RACE技术在cDNA文库中筛选得到TaPrx基因的全长序列并进行生物信息学分析,然后克隆至pET-32a(+),转化E.coliBL21(DE3)后用IPTG进行诱导表... 【目的】克隆TaPrx基因并对其在小麦与条锈菌互作中的功能进行初步分析。【方法】利用PCR方法结合RACE技术在cDNA文库中筛选得到TaPrx基因的全长序列并进行生物信息学分析,然后克隆至pET-32a(+),转化E.coliBL21(DE3)后用IPTG进行诱导表达。通过Real-timeRT-PCR进行表达模式分析。【结果】得到TaPrx基因的全长序列688bp,ORF489bp,编码162个氨基酸残基,分子量17.36kD,等电点5.32,含一个保守的半胱氨酸残基(Cys),不含信号肽及跨膜结构域,亚细胞定位94%的可能性在细胞质。TaPrx融合蛋白分子量38kD,最佳IPTG诱导浓度0.05mmol·L-1,20℃诱导20h可得到最大量的融合蛋白。Real-timeRT-PCR分析表明TaPrx基因在小麦与条锈菌的亲和与非亲和互作中均受诱导表达,分别在接种后24h、18h达到表达高峰。【结论】获得了TaPrx基因特异性的多克隆抗体;TaPrx基因受条锈菌诱导表达,可能参与了小麦与条锈菌互作。但是否参与了小麦受条锈菌侵染后产生的ROS的清除与调节仍需进一步验证。 展开更多
关键词 小麦 条锈菌 TaPrx 克隆 原核表达 REAL-time RT-PCR
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Identification of Sheep Endogenous Beta-Retroviruses with Uterus-Specific Expression in the Pregnant Mongolian Ewe 被引量:6
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作者 QI Jing-wei XU Meng-jie +4 位作者 LIU Shu-ying ZHANG Yu-fei LIU Yue ZHANG Ya-kun CAO Gui-fang 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2013年第5期884-891,共8页
The sheep genome harbours approximately 20 copies of endogenous beta-retroviruses (enJSRVs), and circumstantial evidence suggests that enJSRVs might play a role in mammalian reproduction, particularly placental morp... The sheep genome harbours approximately 20 copies of endogenous beta-retroviruses (enJSRVs), and circumstantial evidence suggests that enJSRVs might play a role in mammalian reproduction, particularly placental morphogenesis. This study was aimed to assess the expression of mRNAs of an enJSRV and its receptor, HYAL2, in the uterus and conceptuses of Mongolian ewes throughout gestation, using real-time reverse transcription polymerase chain reaction and in situ hybridization analysis. The results showed that enJSRV and HYAL2 mRNAs were found to be expressed throughout gestation in the endometrium, chorion, placenta, and conceptus. The enJSRV mRNA was most abundant in the placenta on day 90 of pregnancy, in the endometrium on day 30 and 50, and in the chorion on day 70 and 110. However, HYAL2 mRNA was most abundant in the endometrium on day 30. These differences were all significantly different from each other (P〈0.01). In situ hybridization showed that enJSRV and HYAL2 mRNAs were specifically expressed in endometrial luminal epithelium and glandular epithelium, trophoblastic giant binucleated cells (BNCs), endometrial caruncles, placental cotyledons, stroma, trophectoderm, as well as multinucleated syncytia of the placenta and blood vessel endothelial cells. Collectively, little is known about the molecular mechanisms by which trophoblastic differentiation and multinucleated syncytia formation are regulated by enJSRVs. However, the temporal and spatial distributions of enJSRV expression in the uterus and conceptus indicate that differentiation of BNCs and the formation of a multinucleated syncytiotrophoblast involve enJSRV and possibly its cellular receptor, HYAL2. Therefore, enJSRV and HYAL2 appear to play important roles in the female reproductive physiology in this breed of sheep. 展开更多
关键词 ENJSRV HYAL2 expression real-time reverse transcription polymerase chain reaction in situ hybridizationhybridization Mongolian ewe
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The rhythmic expression of genes controlling flowering time in southern and northern populations of invasive Ambrosia artemisiifolia 被引量:7
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作者 Xiao-Meng Li Da-Yong Zhang Wan-Jin Liao 《Journal of Plant Ecology》 SCIE 2015年第2期207-212,共6页
Aims Flowering time has been suggested to be an important adaptive trait during the dispersal of invasive species,and identifying the molecu-lar mechanisms underlying flowering time may provide insight into the local ... Aims Flowering time has been suggested to be an important adaptive trait during the dispersal of invasive species,and identifying the molecu-lar mechanisms underlying flowering time may provide insight into the local adaptation during the process of invasion.Here,we con-ducted a preliminary exploration on the genetic basis of the differ-entiation of flowering time in Ambrosia artemisiifolia.Methods using relative real-time fluorescent quantitative polymerase chain reaction,we investigated the expression levels of eight flowering-related genes,including AP1,FT,SOC1,CRY2,FKF1,GI,CO2 and SPY,in leaves and flowers at different time points in individuals from northern beijing and southern Wuhan populations that exhibit significant differences in flowering times to identify any rhythmic changes in gene expression and their association with differential flowering times.Important Findings The differentiation of flowering time in the A.artemisiifolia popula-tions was closely associated with five genes involved in flowering pathways.The floral pathway integrators FT and SOC1 and floral meristem identity gene AP1 exhibited increased expression during flowering.The photoreceptor CRY2 in the light-dependent path-way and the SPY gene in the gibberellin pathway displayed specific expression patterns over time.in earlier-flowering beijing plants,CRY2 expression was lower and SPY expression was higher than in Wuhan plants.The expression patterns of these five genes sug-gest a molecular basis for the differentiation of flowering time in A.artemisiifolia. 展开更多
关键词 Ambrosia artemisiifolia flowering time adaptation flowering pathway gene expression
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溴氰菊酯对飞蝗羧酸酯酶基因表达的影响 被引量:6
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作者 张建琴 王燕 +3 位作者 李大琪 杨美玲 张建珍 马恩波 《应用昆虫学报》 CAS CSCD 北大核心 2014年第2期418-424,共7页
【目的】研究溴氰菊酯作用下飞蝗羧酸酯酶基因的mRNA表达特性,为溴氰菊酯的代谢解毒及飞蝗Locusta migratoria防治中抗性风险的评估提供基础资料。【方法】本文采用不同剂量溴氰菊酯处理3龄飞蝗,提取总RNA,体外反转录合成cDNA模板,采用R... 【目的】研究溴氰菊酯作用下飞蝗羧酸酯酶基因的mRNA表达特性,为溴氰菊酯的代谢解毒及飞蝗Locusta migratoria防治中抗性风险的评估提供基础资料。【方法】本文采用不同剂量溴氰菊酯处理3龄飞蝗,提取总RNA,体外反转录合成cDNA模板,采用Real-time PCR技术分析飞蝗羧酸酯酶基因在溴氰菊酯不同浓度和不同时间处理后的表达模式。【结果】飞蝗经不同浓度溴氰菊酯处理12 h后,LmCesA3和LmCesE1表现为诱导效应;除LmCesA2外,其余羧酸酯酶基因经溴氰菊酯LD30剂量处理后分别在不同的时间点表现为诱导效应。【结论】5个羧酸酯酶基因LmCesA1、LmCesA3、LmCesD1、LmCesE1和LmCesI1可以被溴氰菊酯诱导,表明其可能参与飞蝗对溴氰菊酯的代谢解毒及抗性产生。 展开更多
关键词 飞蝗 溴氰菊酯 羧酸酯酶 基因表达 REAL-time PCR
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Recombinant scorpion insectotoxin AaIT kills specifically insect cells but not human cells 被引量:5
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作者 SHENG JIAN JI, FENG LIU, ER Qiu LI, Yu XIAN ZHUThe National Laboratory of Protein Engineering and Plant Genetic Engineering, College of Life Sciences, Peking University, Beijing 100871, China 《Cell Research》 SCIE CAS CSCD 2002年第2期143-150,共8页
The nucleotide sequence deduced from the amino acid sequence of the scorpion insectotoxin AaIT was chemically synthesized and was expressed in Escherichia coli. The authenticity of this in vitro expressed peptide was ... The nucleotide sequence deduced from the amino acid sequence of the scorpion insectotoxin AaIT was chemically synthesized and was expressed in Escherichia coli. The authenticity of this in vitro expressed peptide was confirmed by N-terminal peptide sequencing. Two groups of bioassays, artificial diet incorporation assay and contact insecticidal effect assay, were carried out separately to verify the toxicity of this recombinant toxin. At the end of a 24 h experimental period, more than 60% of the testing diamondback moth (Plutella xylostella) larvae were killed in both groups with LC50 value of 18.4 microM and 0.70 microM respectively. Cytotoxicity assay using cultured Sf9 insect cells and MCF-7 human cells demonstrated that the toxin AaIT had specific toxicity against insect cells but not human cells. Only 0.13 microM recombinant toxin was needed to kill 50% of cultured insect cells while as much as 1.3 microM toxin had absolutely no effect on human cells. Insect cells produced obvious intrusions from their plasma membrane before broken up. We infer that toxin AaIT bind to a putative sodium channel in these insect cells and open the channel persistently, which would result in Na+ influx and finally cause destruction of insect cells. 展开更多
关键词 Amino Acid Sequence Animals Base Sequence Biological Assay Cell Line Cloning Molecular Dose-Response Relationship Drug Electrophoresis Polyacrylamide Gel Escherichia coli Humans Inhibitory Concentration 50 Insects Molecular Sequence Data Peptides Protein Structure Tertiary Recombinant Proteins Research Support Non-U.S. Gov't Scorpion Venoms Sequence Analysis Protein Sodium time Factors Tumor Cells Cultured
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钠/碘转运体在甲状腺癌中的表达 被引量:5
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作者 王志峰 刘勤江 廖世奇 《肿瘤》 CAS CSCD 北大核心 2010年第11期939-943,共5页
目的:探讨钠/碘转运体(sodium iodide symporter,NIS)在甲状腺癌组织中的表达。方法:应用实时荧光定量PCR(real-time fluorescence quantitative-PCR,RFQ-PCR)和免疫组织化学法检测NIS mRNA和蛋白在18例正常甲状腺组织、27例结节性甲状... 目的:探讨钠/碘转运体(sodium iodide symporter,NIS)在甲状腺癌组织中的表达。方法:应用实时荧光定量PCR(real-time fluorescence quantitative-PCR,RFQ-PCR)和免疫组织化学法检测NIS mRNA和蛋白在18例正常甲状腺组织、27例结节性甲状腺肿组织和23例甲状腺癌组织中的表达。结果:甲状腺癌组织中NIS mRNA的表达显著低于正常甲状腺组织和结节性甲状腺肿组织(P=0.002)。免疫组织化学检测结果表明,甲状腺癌组织中NIS蛋白主要定位于细胞质,强阳性表达13例(56.6%)、阳性表达3例(13.0%),强阳性表达率显著高于正常甲状腺组织(P=0.010)。与正常甲状腺组织相比,甲状腺癌组织中NIS mRNA表达下降并且NIS蛋白表达升高者17例(74%),显著高于结节性甲状腺肿组织(χ2=4.428,P=0.035)。结论:甲状腺癌组织中NIS mRNA的表达下降而NIS蛋白的表达增强,推测NIS蛋白主要位于细胞质可能是NIS不能发挥正常的运输功能,导致甲状腺癌患者不能聚碘的重要原因之一。 展开更多
关键词 碘转运体 甲状腺癌 阳性表达率 THYROID carcinoma 甲状腺组织 结节性甲状腺肿 癌组织 NIS蛋白 免疫组织化学法 表达下降 mRNA 细胞质 实时荧光定量 fluorescence real-time PCR 运输功能 检测结果 表达升高 癌患者
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Expression profiles of penaeidin from Fenneropenaeus chinensis in response to WSSV and vibrio infection by real-time PCR 被引量:3
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作者 DONGBo LIUFengsong +2 位作者 XIANGJianhai LIFuhua GAOHongwei 《Acta Oceanologica Sinica》 SCIE CAS CSCD 2005年第2期131-140,共10页
Penaeidin from Chinese shrimp (Fenneropenaeus chinensis) has proved to be one of the most important antimicrobial peptides in the bodies of animals. The relative quantitative real-time PCR method is developed to study... Penaeidin from Chinese shrimp (Fenneropenaeus chinensis) has proved to be one of the most important antimicrobial peptides in the bodies of animals. The relative quantitative real-time PCR method is developed to study through time, the mRNA expression profile of penaeidin in the muscle and haemocyte tissue of Chinese shrimp infected with vibrio (Vibrio anguillarum) and WSSV (white spot syndrome virus). Research results showed that the same pathogens infection experiments produced similar gene expression profile in different tissues while different expression profiles appeared in the same tissues infected by different exterior pathogens. In vibrio infection experiments, a 'U' like expression profile resulted. Expression levels of penaeidin increased and surpassed the non-stimulated level, indicating that penaeidin from Chinese shrimp has noticeable antimicrobial activities. In WSSV infection experiments, the expression profile appeared as an inverse 'U' with the expression of penaeidin gradually decreasing to below baseline level after 24 h. The expression of antimicrobial peptides gene in mRNA level in response to virus infection in shrimp showed that international mechanisms of virus to haemocytes and microbial to haemocytes are completely different. Decline of penaeidins expression levels may be due to haemocytes being destroyed by WSSV or that the virus can inhibit the expression of penaeidins by yet undiscovered modes. The expression profiles of penaeidin in response to exterior pathogen and the difference of expression profiles between vibrio and WSSV infection provided some clues to further understanding the complex innate immune mechanism in shrimp. 展开更多
关键词 penaeidin Fenneropenaeus chinensis expression profile WSSV Vibrio anguillarum real-time PCR
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Revisiting processing time for metaphorical expressions: An eye-tracking study on eye-voice span during sight translation 被引量:4
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作者 Zheng Binghan Zhou Hao 《外语教学与研究》 CSSCI 北大核心 2018年第5期744-759,共16页
This paper aims to investigate the eye-voice span (EVS), the distance between eyeandvoice,insighttranslatingmetaphoricalexpressions(MEs).24MAtranslation students, with no professional translation or interpreting exper... This paper aims to investigate the eye-voice span (EVS), the distance between eyeandvoice,insighttranslatingmetaphoricalexpressions(MEs).24MAtranslation students, with no professional translation or interpreting experience, were asked to conduct a sight translation (STR) task, and the processes were registered by eye-tracker and audio recorder. The qualified eye-tracking and audio data were further analysed by Tobii Studio and Audacity audio processing software. Our findings suggest that the time of the pause preceding an ME was largely, but not entirely, spent in processing the ensuing ME. However, due to the general existence of reading ahead activities in STR, the planning step for sight translating an ME takes place prior to the preceding pause; moreover, due to local processing difficulty causedbytheME,thetimeforreadingaheadintoME(temporalEVS)ismostlygreater than for reading ahead beyond ME. Our findings also reveal that the rate of methodological deviation(causedbythetwodifferentcalculationapproaches)forMEprocessingtimeis around10%,butthetwoprocessingtimeshavedemonstratednostatisticallysignificant difference, validating the processing time calculated by audio data in Zheng&Xiang (2013).We conclude this paper with some reservation on eye-tracking translation research:though powerful in providing solid and informative process data, it has some limitations in clearly probing into intricate human cognitive process. 展开更多
关键词 SIGHT TRANSLATION METAPHORICAL expression eye-voice SPAN processing time EYE-TRACKING
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Molecular cloning and mRNA expression analysis of myosin heavy chain(MyHC)from fast skeletal muscle of grass carp,Ctenopharyngodon idella 被引量:5
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作者 褚武英 符贵红 +6 位作者 宾石玉 蒙涛 周瑞雪 成嘉 赵发兰 张红芳 张建社 《Chinese Journal of Oceanology and Limnology》 SCIE CAS CSCD 2010年第2期239-247,共9页
The myosin heavy chain(MyHC)is one of the major structural and contracting proteins of muscle.We have isolated the cDNA clone encoding MyHC of the grass carp,Ctenopharyngodon idella. The sequence comprises 5 934 bp,in... The myosin heavy chain(MyHC)is one of the major structural and contracting proteins of muscle.We have isolated the cDNA clone encoding MyHC of the grass carp,Ctenopharyngodon idella. The sequence comprises 5 934 bp,including a 5 814 bp open reading frame encoding an amino acid sequence of 1 937 residues.The deduced amino acid sequence showed 69%homology to rabbit fast skeletal MyHC and 73%–76%homology to the MyHCs from the mandarin fish,walleye pollack,white croaker,chum salmon,and carp.The putative sequences of subfragment-1 and the light meromyosin region showed 61.4%–80%homology to the corresponding regions of other fish MyHCs.The tissue-specific and developmental stage-specific expressions of the MyHC gene were analyzed by quantitative real-time PCR.The MyHC gene showed the highest expression in the muscles compared with the kidney,spleen and intestine.Developmentally,there was a gradual increase in MyHC mRNA expression from the neural formation stage to the tail bud stage.The highest expression was detected in hatching larva.Our work on the MyHC gene from the grass carp has provided useful information for fish molecular biology and fish genomics. 展开更多
关键词 grass carp real-time PCR myosin heavy chain fast skeletal muscle gene expression
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