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negative regulation region
相关语句
  负调控区
     Probable Existence of a Negative Regulation Region Within the 3′UTR of Human Phosphoinositide 3-Kinaseγ mRNA
     PI3KγmRNA 3′非翻译区可能存在基因表达负调控区
短句来源
     These results indicate that there is probably a negative regulation region within the 3′AU rich region of PI3Kγ mRNA 3′UTR.
     实验结果提示 ,PI3Kγ基因 3′非翻译区AU富含区内可能存在转录后水平的基因表达负调控区 ,该负调控区可在一定程度上加速mRNA的衰变
短句来源
  相似匹配句对
     On the Proving Regulation about Negative Fact
     论消极事实的证明规则
短句来源
     Negative regulation of homocysteine metabolism by stress in rats
     应激对同型半胱氨酸代谢的负性调节
短句来源
     Regulation and Control
     约束与控制
短句来源
     Regulation and Corruption
     政府规制与腐败
短句来源
     starch negative;
     革兰氏染色阴性;
短句来源
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FHL2, a member of LIM only protein family, plays an important role in transcription regulation, apoptosis, cancer development and progression. In this study, a mammalian transcription activation system was constructed by using DNA binding domain(DBD) of GAL4 and luciferase reporter gene with DBD binding sequence, and used for mapping of FHL2 transcription activation domain. First, the coding sequence of GAL4 DBD was inserted into expression vector pcDNA3, generating the pDBD recombinant plasmid, then the wild...

FHL2, a member of LIM only protein family, plays an important role in transcription regulation, apoptosis, cancer development and progression. In this study, a mammalian transcription activation system was constructed by using DNA binding domain(DBD) of GAL4 and luciferase reporter gene with DBD binding sequence, and used for mapping of FHL2 transcription activation domain. First, the coding sequence of GAL4 DBD was inserted into expression vector pcDNA3, generating the pDBD recombinant plasmid, then the wild type FHL2 and its mutants were fused in frame with GAL4 DBD, resulting in expression vectors for wild type FHL2 and its mutants. All of the recombinant plasmids were transfected into 293T cells. Western blot assay showed that all of the fusion proteins were expressed. The analysis of FHL2 transcription activation properties by using the GAL4 luciferase reporter gene indicated that wild type FHL2 had activation activity in both 293T and MCF 7 cells. The deletion of the half LIM domain at the N terminus severely impaired the capacity of FHL2 to stimulate transcription. The mutant lacking the LIM domain at the C terminus was totally inactive, while the deletion of two LIM domains at the C terminus partially recovered its ability to stimulate transcription. The deletion of the second LIM domain at C terminus did not alter the activation capacity of FHL2. These results suggest that the last LIM domain at the C terminus of FHL2 is critical for its transcription activation function, the second LIM domain at the C terminus may be a negative regulation region, but this negative regulation depends on the last LIM domain. Mapping of transcription activation domain of FHL2 lays solid basis for further study on various FHL2 functions.

LIM蛋白家族成员FHL2 (fourandhalfLIMdomainprotein)在转录调节、细胞凋亡及肿瘤的发生发展中都起着重要作用。利用GAL4转录因子中的DNA结合结构域 (DBD)和含有与DBD结合序列的荧光素酶报告基因(GAL4 LUC)构建了哺乳动物细胞转录激活系统 ,并利用该系统定位了FHL2的转录激活结构域。首先将GAL4 DBD序列以正确读框插入到pcDNA3载体的多克隆位点中 ,构建成真核表达载体pDBD ,再将野生型FHL2及其不同片段以正确读框与pDBD中GAL4 DBD序列融合 ,构建成野生型FHL2及其缺失突变体表达载体。将这些表达载体分别瞬时转染 2 93T胚胎肾细胞 ,野生型FHL2及其缺失突变体都得到了表达。利用GAL4 荧光素酶报告基因对野生型FHL2及其不同突变体的转录激活活性检测表明 ,在 2 93T胚胎肾细胞和乳腺癌MCF 7细胞中 ,野生型FHL2具有转录激活活性 ,缺失N端半个LIM结构域使FHL2转录激活活性降低 ,缺失C末端第二个LIM结构域对FHL2的转录激活功能影响不大 ,缺失C末端最后一个LIM结构域则使FHL2的转录激活功能完全丧失 ,而C...

LIM蛋白家族成员FHL2 (fourandhalfLIMdomainprotein)在转录调节、细胞凋亡及肿瘤的发生发展中都起着重要作用。利用GAL4转录因子中的DNA结合结构域 (DBD)和含有与DBD结合序列的荧光素酶报告基因(GAL4 LUC)构建了哺乳动物细胞转录激活系统 ,并利用该系统定位了FHL2的转录激活结构域。首先将GAL4 DBD序列以正确读框插入到pcDNA3载体的多克隆位点中 ,构建成真核表达载体pDBD ,再将野生型FHL2及其不同片段以正确读框与pDBD中GAL4 DBD序列融合 ,构建成野生型FHL2及其缺失突变体表达载体。将这些表达载体分别瞬时转染 2 93T胚胎肾细胞 ,野生型FHL2及其缺失突变体都得到了表达。利用GAL4 荧光素酶报告基因对野生型FHL2及其不同突变体的转录激活活性检测表明 ,在 2 93T胚胎肾细胞和乳腺癌MCF 7细胞中 ,野生型FHL2具有转录激活活性 ,缺失N端半个LIM结构域使FHL2转录激活活性降低 ,缺失C末端第二个LIM结构域对FHL2的转录激活功能影响不大 ,缺失C末端最后一个LIM结构域则使FHL2的转录激活功能完全丧失 ,而C末端缺失 2个LIM结构域使FHL2转录激活活性又有所恢复。这说明FHL2C末端最后一个LIM结构域对其转录激活功能是必需的 ,而C末端第二个LIM结构域可能对FHL2的转录激活功能有负调控作用 ,这种负调控作用取决于

In order to study whether there is a functional regulation region within the 3′untranslational region (UTR) of human phosphoinositide 3 kinaseγ(PI3Kγ)mRNA, the 3′UTR was analyzed through bioinformatics. An AU rich region (AUR)of about 0 9 kb was found. Within the AU rich region there are 4 AU rich elements (AREs) and a region which is homologous with many other UTRs. The AU rich region was inserted into the 3′UTR of egfp reporter gene, and pcDNA3 egfp AUR expressing vector was constructed. The...

In order to study whether there is a functional regulation region within the 3′untranslational region (UTR) of human phosphoinositide 3 kinaseγ(PI3Kγ)mRNA, the 3′UTR was analyzed through bioinformatics. An AU rich region (AUR)of about 0 9 kb was found. Within the AU rich region there are 4 AU rich elements (AREs) and a region which is homologous with many other UTRs. The AU rich region was inserted into the 3′UTR of egfp reporter gene, and pcDNA3 egfp AUR expressing vector was constructed. The expression of egfp containing the 3′UTR of PI3Kγ was significantly reduced by 2~3 times in NIH 3T3, 7402 and K562 cells. After treated with actinomycin D (5 μg/ml) total RNA from stably transfected K562 cells were successively extracted with an interval of 2 hours, and the stability of egfp mRNA was analyzed through Northern blotting. The result showed that the AU rich region of PI3Kγ could, to some extent, accelerate the decay of egfp mRNA. These results indicate that there is probably a negative regulation region within the 3′AU rich region of PI3Kγ mRNA 3′UTR.

为探索人磷脂酰肌醇 3 激酶γ(phosphoinositide 3 kinasePI3Kγ)基因 3′端非翻译区内AU富含区是否在基因表达调控中起作用 ,首先通过生物信息学分析发现在其 3′端非翻译区 (UTR)内存在0 9kb的AU富含区 ,其中包括 4个AU富含元件 ,以及 1个与众多基因非翻译区高度同源的长 130个碱基的区域 .将AU富含区插入报告基因egfp的下游构建pcDNA3 egfp AUR表达载体 .将表达载体转导NIH 3T3,74 0 2及K5 6 2细胞 ,流式细胞检测egfp的表达情况 .PI3Kγ基因 3′非翻译区AU富含区可显著降低egfp的表达 2~ 3倍 (P <0 0 1) .利用放线菌素D阻断RNA转录后 ,Northern印迹分析结果显示egfp AURmRNA较egfpmRNA不稳定 .实验结果提示 ,PI3Kγ基因 3′非翻译区AU富含区内可能存在转录后水平的基因表达负调控区 ,该负调控区可在一定程度上加速mRNA的衰变

Objective: To identify a binding protein to a 30 bp-negative regulation region upstream of NKX 3.1 gene. Methods: The 30 bp-sequence was synthesized and labeled with digoxigenin by terminal transferase. The nucleic extracts were isolated and bound to the labeled probe of 30bp-sequence. Electrophoresis mobility shift assay (EMSA) was used to identify the binding proteins to the 30 bp-sequence. Results: The proteins that bound specifically to 30 bp-sequence were found in the nucleic extracts of LNCaP cell...

Objective: To identify a binding protein to a 30 bp-negative regulation region upstream of NKX 3.1 gene. Methods: The 30 bp-sequence was synthesized and labeled with digoxigenin by terminal transferase. The nucleic extracts were isolated and bound to the labeled probe of 30bp-sequence. Electrophoresis mobility shift assay (EMSA) was used to identify the binding proteins to the 30 bp-sequence. Results: The proteins that bound specifically to 30 bp-sequence were found in the nucleic extracts of LNCaP cell line and other tested tumor cell lines. However, the binding proteins to 30 bp-sequence were different in various tested cell lines. Conclusions: The event of 30 bp-sequence-involved negative regulation for NKX 3.1 gene is common in different cell lines. Different binding proteins and mechanisms may be involved in the regulation of NKX 3.1 gene in different cells.

目的:鉴定NKX3.1基因上游30bp负调控区的结合蛋白。方法:人工合成30bp负调控序列,用末端转移酶对其进行地高辛标记;提取细胞核蛋白,采用电泳迁移率变动分析(EMSA)方法,检测细胞核提取液中与30bp负调控序列特异结合的核蛋白。结果:在LNCaP细胞及其他不同肿瘤细胞系核提取液中,检测到与30bp负调控序列特异结合的核蛋白,但在不同的细胞系中有不同种类的结合蛋白。结论:NKX3.1基因上游的30bp负调控机制在不同的细胞中普遍存在,不同的细胞核中有不同的结合蛋白,可能涉及不同的调控机理。

 
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