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截断突变体
相关语句
  truncation mutant
     Methods Truncation mutant(Arg832 Arg879)of band 3 C terminus was amplified from pFAST Bac AE1 C end recombinant plasmid with polymerase chain reaction(PCR). The cDNA fragment of AE1 C end truncation mutant was inserted into the yeast two hybrid activation domain vector.
     方法 采用PCR方法 ,从pFAST Bac AE1 C end杆状病毒表达载体中扩增出AE1 C end截断突变体 (Arg832 Arg879) ,将突变体的cDNA片段插入酵母双杂交系统GAL4AD端表达载体中 ,观察其在选择性培养基上的表达情况。
短句来源
     Results We successfully constructed the truncation mutant of AE1 C end and identified that there was more than one site of direct interaction between the AE1 C end truncation mutant and GPA.
     结果 成功构建了AE1 C 末端截断突变体酵母双杂交的AD端表达质粒 ,证实带 3蛋白C 末端截断突变体与GPA不止有一个相互作用位点。
短句来源
  truncated mutants
     Methods The plasmids which can express the truncated mutants of the TPR tandem clusters at different region of CRLP were constructed, stable transfected BEL7402 cell lines were established.
     方法 构建了不同TPR串联簇的截断突变体的表达质粒后 ,建立了稳定转染的BEL740 2细胞株。
短句来源
     The neuron differentiation function of different truncated mutants was observed by PC12 transient transfection assay.
     通过瞬时转染PC12细胞实验研究不同截断突变体的促神经细胞分化功能。
短句来源
  相似匹配句对
     The cutting problem
     截断切割
短句来源
     LIGHT RESPOSIVE MUTANTS IN PLANTS
     植物光反应突变体
短句来源
     Mutants of Subtilisin E
     枯草蛋白酶E的突变体
短句来源
     ON THE ASYMPTOTIC DISTRIBUTION OF HEAVILY TRIMMED SUMS
     重截断和的渐近分布
短句来源
     The neuron differentiation function of different truncated mutants was observed by PC12 transient transfection assay.
     通过瞬时转染PC12细胞实验研究不同截断突变体的促神经细胞分化功能。
短句来源
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  truncation mutant
To delineate the role of the Cldn cytoplasmic tail domain in epidermal differentiation, we engineered transgenic mice targeting the overexpression of a Cldn6 cytoplasmic tail-truncation mutant in the epidermis.
      
We have compared the functional and structural integrity of gap junction channels assembled from a Cx45 truncation mutant with those of gap junction channels assembled from wild-type (wt) Cx45 and Cx43.
      
The truncation mutant lacks the last 26 amino acids of the COOH-terminus, including nine serine phosphorylation sites that are associated with regulatory processes of these channels.
      
Beyond this, the modified toxin, or a truncation mutant of the toxin, may have utility as a carrier in the construction of other oral vaccines.
      
A complete NMR spectral assignment of the lipid-free mouse apolipoprotein A-I (apoAI) C-terminal truncation mutant, apoAI(1-216)
      
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  truncated mutants
A series of truncated mutants of NifA were constructed to determine the structural elements at the central domain critical for multimerization.
      
MAL63/mal64 fusions of the truncated mutants suggest that residues in the N-terminal 100 residues containing the DNA-binding domain also modulate basal expression.
      
The antigenic epitope recognized by this mAb was mapped within the residues 1-152 of EBV DNase by reacting the mAb with three distinct truncated mutants.
      
Analysis of the proteins by1H-NMR showed that the truncated mutants have higher exchange rates with solvent, indicating a less rigid structure.
      
Studies on the truncated mutants showed that the intact EF-3 and EF-4 were essential for the binding of VILIP1 with Ca2+ and Mg2+.
      
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Band 3 membrane domain were expressed on yeast membrane surface by pYD1 yeast display system. The expressed membrane domain showed anion transport activity and DIDS could inhibit this function of membrane domain. About 1 500 bp cDNA fragment of truncation mutagenesis of band 3 membrane domain were amplified by PCR, which knockout Ala908~Val911, Asp896~Val911, Lys892~Val911 and Asn880~Val911 of band 3 respectively. After being sequenced, the four gene fragments cloned into Eco RⅠ~ Bam HⅠ sites of pYD1....

Band 3 membrane domain were expressed on yeast membrane surface by pYD1 yeast display system. The expressed membrane domain showed anion transport activity and DIDS could inhibit this function of membrane domain. About 1 500 bp cDNA fragment of truncation mutagenesis of band 3 membrane domain were amplified by PCR, which knockout Ala908~Val911, Asp896~Val911, Lys892~Val911 and Asn880~Val911 of band 3 respectively. After being sequenced, the four gene fragments cloned into Eco RⅠ~ Bam HⅠ sites of pYD1. The recombinant plasmids pYD1 Trunc4/Trunc16/Trunc20/Trunc32 were transformed into yeast EBY100. As control, pYD1 mdb3 was also transformed. After four groups fusion protein were expressed after adding galactose, the Cl - transport activity was measured by using a fluorescent probe SPQ. The result demonstrated that the transport activity of band 3 was decreased when knockout Lys892~Val911 of AE1 C terminal domain, but the transport activity didnt decrease further when knockout Asn880~Val911 of AE1 C terminal domain. These results showed that Lys892~Phe895 amino acids influenced the anion transport of band 3 transmembrane domain.

采用酵母表面展示系统 ,表达带 3蛋白膜段结构域 (Gln4 0 4~Val911)至酵母细胞膜 ,功能研究表明 ,表达后的膜段结构域具有离子转运的活性 ,同时 ,带 3蛋白抑制剂 4 ,4′ 二异硫氰 2 ,2′ 二黄酸芪 (DIDS)能够抑制其离子转运的功能 .利用PCR方法 ,以 pFAST Bac mdb3为模板扩增出带 3蛋白膜段结构域的 4种截断突变体 ,分别去除带 3蛋白C端域后 4个 (Ala90 8~Val911)、 16个 (Asp896~Val911)、 2 0个 (Lys892~Val911)、 32个(Asn880~Val911)氨基酸序列 ,测序后将其克隆至表达载体 pYD1上 ,构建酵母表达质粒 pYD1 Trunc4、pYD1 Trunc16、pYD1 Trunc2 0和 pYD1 Trunc32 ,诱导 4组突变体的蛋白质表达 .然后测定Cl-的转运活性 ,结果发现去除后 2 0个 (Lys892~Val911)氨基酸残基后 ,离子转运活性明显下降 ,而去除后 32个 (Asn880~Val911)后 ,离子转运没有进一步下降 ,说明Lys892~Phe895 4...

采用酵母表面展示系统 ,表达带 3蛋白膜段结构域 (Gln4 0 4~Val911)至酵母细胞膜 ,功能研究表明 ,表达后的膜段结构域具有离子转运的活性 ,同时 ,带 3蛋白抑制剂 4 ,4′ 二异硫氰 2 ,2′ 二黄酸芪 (DIDS)能够抑制其离子转运的功能 .利用PCR方法 ,以 pFAST Bac mdb3为模板扩增出带 3蛋白膜段结构域的 4种截断突变体 ,分别去除带 3蛋白C端域后 4个 (Ala90 8~Val911)、 16个 (Asp896~Val911)、 2 0个 (Lys892~Val911)、 32个(Asn880~Val911)氨基酸序列 ,测序后将其克隆至表达载体 pYD1上 ,构建酵母表达质粒 pYD1 Trunc4、pYD1 Trunc16、pYD1 Trunc2 0和 pYD1 Trunc32 ,诱导 4组突变体的蛋白质表达 .然后测定Cl-的转运活性 ,结果发现去除后 2 0个 (Lys892~Val911)氨基酸残基后 ,离子转运活性明显下降 ,而去除后 32个 (Asn880~Val911)后 ,离子转运没有进一步下降 ,说明Lys892~Phe895 4个氨基酸残基在带 3蛋白的离子转运过程中发挥重要作用

Objective CRLP(Crn-like protein) is a novel gene that has been cloned by author's lab. The protein product of the CRLP has fifteen TPR(tetratricopeptide repeat) motifs, which can be divided into three TPR tandem clusters, named as D1, D2, D3 according to their position on the CRLP molecule. The relationship between the TPR tandem clusters and the function of CRLP is to be explored. Methods The plasmids which can express the truncated mutants of the TPR tandem clusters at different region of CRLP were constructed,...

Objective CRLP(Crn-like protein) is a novel gene that has been cloned by author's lab. The protein product of the CRLP has fifteen TPR(tetratricopeptide repeat) motifs, which can be divided into three TPR tandem clusters, named as D1, D2, D3 according to their position on the CRLP molecule. The relationship between the TPR tandem clusters and the function of CRLP is to be explored. Methods The plasmids which can express the truncated mutants of the TPR tandem clusters at different region of CRLP were constructed, stable transfected BEL7402 cell lines were established. The growth status of stable transfected BEL7402 cells and the changes of cell drug-resistance after adding chemotherapy drug adriamycin or cisplatin in the cell medium were examined by cell growth curve assay. The neuron differentiation function of different truncated mutants was observed by PC12 transient transfection assay. Results The proliferation of the BEL7402 cells stably transfected with the plasmid expressing TPR tandem clusters D2 and D3 were inhibited while the drug-resistance of the cells transfected with the plasmid expressing TPR tandem cluster D1 increased. PC12 transient transfection assay revealed that TPR tandem clusters D2 and D3 could induce PC12 differentiation. Conclusion TPR tandem cluster D1 of CRLP are related to cell drug-resistance. TPR tandem clusters D2 and D3 have the role of inhibiting cell grouth and may induce neuron differentiation.

目的 CRLP(Crn likeprotein)基因是本室克隆的新基因 ,它的蛋白表达产物具有 15个TPR(tetratricopeptiderepeat)基序 ,可按在CRLP分子上的位置把它们分为 3个TPR串联簇 ,分别命名为D1、D2、D3。本研究拟探讨这 3个TPR串联簇与CRLP功能的关系。方法 构建了不同TPR串联簇的截断突变体的表达质粒后 ,建立了稳定转染的BEL740 2细胞株。用细胞生长曲线实验检测了稳定转染细胞株的生长状况及观察在培养液中添加化疗药物阿霉素或顺铂后细胞耐药性的变化 ;通过瞬时转染PC12细胞实验研究不同截断突变体的促神经细胞分化功能。结果 稳定表达TPR串联簇D2、D3的BEL740 2细胞的生长受到了抑制 ,稳定表达TPR串联簇D1的BEL740 2细胞耐药性提高 ,瞬时表达TPR串联簇D2、D3的PC12细胞有分化现象。结论 CRLP分子的TPR串联簇D1可提高细胞耐药性 ,TPR串联簇D2和D3有抑制细胞生长的功能 ,并可促神经细胞分化。

Objective To investigate the interaction sites of band 3 C terminus with glycophorin A(GPA).Methods Truncation mutant(Arg832 Arg879)of band 3 C terminus was amplified from pFAST Bac AE1 C end recombinant plasmid with polymerase chain reaction(PCR). The cDNA fragment of AE1 C end truncation mutant was inserted into the yeast two hybrid activation domain vector. The recombinant plasmid was co transformed along with pGBKT7 GPA binding domain vector into yeast strain AH109 for nutrition deficient...

Objective To investigate the interaction sites of band 3 C terminus with glycophorin A(GPA).Methods Truncation mutant(Arg832 Arg879)of band 3 C terminus was amplified from pFAST Bac AE1 C end recombinant plasmid with polymerase chain reaction(PCR). The cDNA fragment of AE1 C end truncation mutant was inserted into the yeast two hybrid activation domain vector. The recombinant plasmid was co transformed along with pGBKT7 GPA binding domain vector into yeast strain AH109 for nutrition deficient and β galactosidase activity analysis.Results We successfully constructed the truncation mutant of AE1 C end and identified that there was more than one site of direct interaction between the AE1 C end truncation mutant and GPA.Conclusion The sites of interaction of Band 3 with GPA are not only located in the 32 amino acid of C terminus but also lied within the last two transmembrane domain of band 3.

目的 利用酵母双杂交系统探寻带 3蛋白与血型糖蛋白A(GPA)的相互作用位点。方法 采用PCR方法 ,从pFAST Bac AE1 C end杆状病毒表达载体中扩增出AE1 C end截断突变体 (Arg832 Arg879) ,将突变体的cDNA片段插入酵母双杂交系统GAL4AD端表达载体中 ,观察其在选择性培养基上的表达情况。用醋酸锂法将构建好的重组质粒与BD端表达质粒共同转化酵母菌AH10 9,经酵母双杂交营养缺陷培养和 β 半乳糖苷酶检测证实带 3蛋白C 末端与GPA之间的作用位点。结果 成功构建了AE1 C 末端截断突变体酵母双杂交的AD端表达质粒 ,证实带 3蛋白C 末端截断突变体与GPA不止有一个相互作用位点。结论 带 3蛋白与GPA的作用位点不仅在带 3蛋白的酸性C端域 ,还涉及其最后两次跨膜域

 
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