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91.
92.
Xia S Wang M Lee HR Sinha A Blaha G Christian T Wang J Konigsberg W 《Journal of molecular biology》2011,406(4):558-10149
We have previously observed that stepwise replacement of amino acid residues in the nascent base-pair binding pocket of RB69 DNA polymerase (RB69pol) with Ala or Gly expanded the space in this pocket, resulting in a progressive increase in misincorporation. However, in vivo results with similar RB69pol nascent base-pair binding pocket mutants showed that mutation rates, as determined by the T4 phage rI forward assay and rII reversion assay, were significantly lower for the RB69pol S565G/Y567A double mutant than for the Y567A single mutant, the opposite of what we would have predicted. To investigate the reasons for this unexpected result, we have determined the pre-steady-state kinetic parameters and crystal structures of relevant ternary complexes. We found that the S565G/Y567A mutant generally had greater base selectivity than the Y567A mutant and that the kinetic parameters for dNMP insertion, excision of the 3′-terminal nucleotide residue, and primer extension beyond a mispair differed not only between these two mutants but also between the two highly mutable sequences in the T4 rI complementary strand. Comparison of the crystal structures of these two mutants with correct and incorrect incoming dNTPs provides insight into the unexpected increase in the fidelity of the S565G/Y567A double mutant. Taken together, the kinetic and structural results provide a basis for integrating and interpreting in vivo and in vitro observations. 相似文献
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酪氨酸羟化酶(tyrosine hydroxylase,TH)是儿茶酚胺(catech01amines,CAs)合成过程中的限速酶,提高或抑制该酶的活性可大幅度影响CAs的合成。近年来对于TH的关注越来越多,大量的研究表明,TH含量及活性的异常改变能通过影响CAs的含量而导致相应生理功能的异常,从而导致疾病。本文将TH的结构功能、分布、表达变化及其与视网膜疾病关系做一综述。 相似文献
95.
取食转Bt基因棉花上的棉蚜对丽草蛉发育和繁殖的影响 总被引:6,自引:0,他引:6
为明确转Bt基因抗虫棉对捕食性天敌的生态效应 ,比较了 2种转Bt基因抗虫棉 (GK-1 2和新棉99B)和 1种常规棉上的棉蚜Chrysopaformosa对丽草蛉Aphisgossypii发育和繁殖的影响。结果表明 ,(1 )与取食常规棉泗棉 3号上棉蚜的对照相比 ,取食转Bt基因抗虫棉GK- 1 2上棉蚜的第 1代和第 2代丽草蛉 ,其幼虫期和茧期的死亡率、幼虫和茧的发育历期、茧重及成虫性比等均与对照无显著差异 ;但第 1代成虫的产卵量减少了 2 88 0粒 ,与对照差异显著 (P <0 . 0 1 ) ;第 2代成虫所产卵的孵化率为 64. 0 % ,显著低于对照的 77. 7% (P <0 0 1 )。 (2 ) 2个品种的Bt棉对丽草蛉发育和繁殖的影响也略有差异。与取食GK 1 2上棉蚜的个体相比 ,取食新棉 99B上蚜虫的第 1代丽草蛉 ,其幼虫发育历期缩短了 0 . 6d(P <0 0 1 ) ,茧期缩短了 0 . 7d(P <0 . 0 1 ) ,茧重降低了 1 . 2mg(P <0 .0 1 ) ,幼虫期和茧期的死亡率、茧重以及成虫性比等则无显著差异 ;雌成虫产卵前期、产卵期、产卵量和寿命等繁殖学特性也无显著差异 ,但其成虫所产卵的孵化率为 65. 0 % ,显著低于取食GK- 1 2上棉蚜的处理 (72 . 7% ,P <0. 0 1 )。分别取食 2种Bt棉上的棉蚜 ,对第 2代丽草蛉的发育和繁殖的影响差异则较小。 相似文献
96.
人为水土流失是一种人类干扰下的典型景观退化现象.将人为侵蚀在空间上的表现称之为人为侵蚀景观.在遥感和GIS技术支持下,结合大量的实地调查资料,对珠海最主要的人为侵蚀景观类型——土石场开展空间分布分析和景观生态影响评价.调查范围内珠海土石场有235个,其中三灶、井岸两镇(区)分布最为集中,分别为28、29个.缓冲区分析表明,土石场的分布与距交通道路的远近呈对数线性相关,其中在距交通道路100~200m距离范围内分布最为集中.设计景观生态影响度评价指数,对面积大于5000m^2的152个土石场进行景观生态影响度评价结果表明,对周边景观生态环境影响极强的土石场有9个,属于强一级的有17个,应首先对这些土石场开展景观生态恢复. 相似文献
97.
98.
Höng JC Ivanov NV Hodor P Xia M Wei N Blevins R Gerhold D Borodovsky M Liu Y 《Journal of molecular biology》2004,337(2):307-317
We have combined protein motif search and gene finding methods to identify genes encoding proteins containing specific domains. Particularly, we have focused on finding new human genes of the cadherin superfamily proteins, which represent a major group of cell-cell adhesion receptors contributing to embryonic neuronal morphogenesis. Models for three cadherin protein motifs were generated from over 100 already annotated cadherin domains and used to search the complete translated human genome. The genomic sequence regions containing motif "hits" were analyzed by eukaryotic GeneMark.hmm to identify the exon-intron structure of new genes. Three new genes CDH-J, PCDH-J and FAT-J were found. The predicted proteins PCDH-J and FAT-J were classified into protocadherin and FAT-like subfamilies, respectively, based on the number and organization of cadherin domains and presence of subfamily-specific conserved amino acid residues. Expression of FAT-J was shown in almost all tested tissues. The exon-intron organization of CDH-J was experimentally verified by PCR with specifically designed primers and its tissue-specific expression was demonstrated. The described methodology can be applied to discover new genes encoding proteins from families with well-characterized structural and functional domains. 相似文献
99.
Inorganic nanoparticles (NPs) including semiconductor quantum dots (QDs), iron oxide NPs and gold NPs have been developed as contrast agents for diagnostics by molecular imaging. Compared with traditional contrast agents, NPs offer several advantages: their optical and magnetic properties can be tailored by engineering the composition, structure, size and shape; their surfaces can be modified with ligands to target specific biomarkers of disease; the contrast enhancement provided can be equivalent to millions of molecular counterparts; and they can be integrated with a combination of different functions for multimodal imaging. Here, we review recent advances in the development of contrast agents based on inorganic NPs for molecular imaging, and also touch on contrast enhancement, surface modification, tissue targeting, clearance and toxicity. As research efforts intensify, contrast agents based on inorganic NPs that are highly sensitive, target-specific and safe to use are expected to enter clinical applications in the near future. 相似文献
100.