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171.
DNA ligases play a pivotal role in DNA replication, repair and recombination. Reactions catalyzed by DNA ligases consist of three steps: adenylation of the ligase in the presence of ATP or NAD+, transferring the adenylate moiety to the 5'-phosphate of the nicked DNA substrate (deadenylation) and sealing the nick through the formation of a phosphodiester bond. Thermus thermophilus HB8 DNA ligase (Tth DNA ligase) differs from mesophilic ATP-dependent DNA ligases in three ways: (i) it is NAD+ dependent; (ii) its optimal temperature is 65 instead of 37 degrees C; (iii) it has higher fidelity than T4 DNA ligase. In order to understand the structural basis underlying the reaction mechanism of Tth DNA ligase, we performed site-directed mutagenesis studies on nine selected amino acid residues that are highly conserved in bacterial DNA ligases. Examination of these site-specific mutants revealed that: residue K118 plays an essential role in the adenylation step; residue D120 may facilitate the deadenylation step; residues G339 and C433 may be involved in formation of the phosphodiester bond. This evidence indicates that a previously identified KXDG motif for adenylation of eukaryotic DNA ligases [Tomkinson, A.E., Totty, N.F., Ginsburg, M. and Lindahl, T. (1991) Proc. Natl. Acad. Sci. USA, 88, 400-404] is also the adenylation site for NAD+-dependent bacterial DNA ligases. In a companion paper, we demonstrate that mutations at a different Lys residue, K294, may modulate the fidelity of Tth DNA ligase. 相似文献
172.
13C MAS NMR studies of crystalline cholesterol and lipid mixtures modeling atherosclerotic plaques. 下载免费PDF全文
Cholesterol and cholesteryl esters are the predominant lipids of atherosclerotic plaques. To provide fundamental data for the quantitative study of plaque lipids in situ, crystalline cholesterol (CHOL) and CHOL/cholesteryl ester (CE) mixtures with other lipids were studied by solid-state nuclear magnetic resonance with magic-angle-sample spinning. Highly distinctive spectra for three different crystalline structures of CHOL were obtained. When CHOL crystals were mixed with isotropic CE oil, solubilized CHOL (approximately 13 mol % CHOL) was detected by characteristic resonances such as C5, C6, and C3; the excess crystalline CHOL (either anhydrous or monohydrate) remained in its original crystalline structure, without being affected by the coexisting CE. By use of 13C-enriched CHOL, the solubility of CHOL in the CE liquid-crystalline phase (approximately 8 mol %) was measured. When phosphatidylcholine was hydrated in presence of CHOL and CE, magic-angle-sampling nuclear magnetic resonance revealed liquid-crystalline CHOL/phosphatidylcholine multilayers with approximately an equal molar ratio of CHOL/phosphatidylcholine. Excess CHOL existed in the monohydrate crystalline form, and CE in separate oil or crystalline phases, depending on the temperature. The magic-angle-sampling nuclear magnetic resonance protocol for identifying different lipid phases was applied to intact (ex vivo) atherosclerotic plaques of cholesterol-fed rabbits. Liquid, liquid-crystalline, and solid phases of CE were characterized. 相似文献
173.
Hong Luo Benoît Van Coppenolle Marc Seguin Marc Boutry 《Molecular breeding : new strategies in plant improvement》1995,1(1):51-63
Using fourteen random mitochondrial DNA probes, we have examined restriction fragment length polymorphism (RFLP) in wild and cultivatedHevea brasiliensis. A total of 395 accessions, including 345 from various prospectings collected in Brazil, Colombia and Peru and 50 cultivated clones, were analyzed. Two other species (H. benthamiana andH. pauciflora) were also included in the study for comparison. The high level of mitochondrial polymorphism allowed us to divide all the accessions analyzed into 212 distinct genotypes. The genetic variability of cultivated clones was limited to four genotypes forming two clusters. In contrast, considerable genetic variation was found in the wild collections. In almost all cases, accessions displaying the same RFLP profile were restricted to the same geographical area (same or neighbor administrative districts). In addition, accessions whose genetic closeness was predicted by RFLP profiles were also clustered according to geographical origin. In a few cases, however, similar RFLP profiles were found for accessions originating from geographically distant districts. This discrepancy can be explained either by seed dispersion (by river) or possibly by similar genetic events occurring independently in different geographical locations. Chloroplast DNA RFLP was also analyzed in 217 accessions, representative of 126 distinct mitochondrial genotypes. Very few differences were found, indicating that the chloroplast genome is more highly conserved than the mitochondrial genome. 相似文献
174.
176.
中药凌霄花的药理学考察 总被引:2,自引:0,他引:2
实验结果表明,凌霄花对离体猪冠状动脉条具有抑制收缩的作用;对大鼠血栓形成有抑制作用;能加快红细胞电泳,增加红细胞电泳率,使血液红细胞处于分散状态;对离体孕子宫能显著增强收缩活性,增加收缩频率及增强收缩强度。以上结果可初步说明,凌霄花行血去瘀的作用及其“孕妇慎用”的合理性。凌霄花的3种混淆品(白泡桐花,毛泡桐花,兰考泡桐花)的药理作用与凌霄花不同,故不能作凌霄花使用。 相似文献
177.
羊草草地枯枝落叶中N,P,K变化动态 总被引:7,自引:2,他引:5
羊草草地枯枝落叶中N、P、K变化动态郭继勋,祝廷成(东北师范大学国家草地生态工程实验室,长春130024)DynamicsofN,PandKinlittersonAneurolopidiumchinensegrassland.¥GuoJixunand... 相似文献
178.
中药凌霄花的研究进展 总被引:1,自引:0,他引:1
本文对中药凌霄花的研究进展进行了综述,包括生药学研究、化学成分研究、药理学研究及临床应用。 相似文献
179.
180.
本文用免疫细胞化学方法调查了生长抑素(SOM)免疫反应神经元在人胎丘脑网状核内的分布。流产的胚胎3例,胎龄分别为18周,23周,32周。意外死亡足月新生儿1例、在18周胚胎的丘脑网状核内可见少数染色较浅的SOM免疫反应阳性神经元,呈圆形。从18周到32周,SOM免疫反应阳性细胞数明显增多,突起更丰富。在足月新生儿,SOM阳性细胞数较32周有所减少。结果表明,SOM阳性神经元存在于人胎丘脑网状核内,并且有一定的发育过程。出现于人脑发育的早期阶段,可能在中枢神经系统的发育过程中起重要的作用。 相似文献