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991.
We describe a new approach to in vitro DNA recombination termed Separate-Mixing method in this study. The reaction process of this method consists of two stages: at the first stage the reaction was implemented in two parallel teams, which generated random recombination by template-switching of growing polynucleotides from primers in the presence of unidirectional single-stranded DNA fragments used as templates, and then both teams were mixed together for further extension and recombination of DNA sequences at the second stage. Because of the particular strategy, the reaction process was also accompanied by the other two processes of DNA shuffling and StEP simultaneously. Two AdoMet synthetase genes sam2 from Saccharomyces cerevisiae and metK from Escherichia coli, which have only 56% homology on the DNA level were used for recombination with Separate-Mixing method. DNA recombination was available after a single round of reaction. With sequencing of 10 randomly selected recombinants, no unshuffled parental clone was found, and also no unexpected insertion, deletion or rearrangement was detected. An evolved gene sam' was obtained after screen and selection, which could obviously increase the accumulation of AdoMet in S. cerevisiae. 相似文献
992.
Lin Q Xiu Y Jiang Y Tsurui H Nakamura K Kodera S Ohtsuji M Ohtsuji N Shiroiwa W Tsukamoto K Amano H Amano E Kinoshita K Sudo K Nishimura H Izui S Shirai T Hirose S 《Journal of immunology (Baltimore, Md. : 1950)》2006,177(3):1646-1654
Immune complex (IC)-mediated tissue inflammation is controlled by stimulatory and inhibitory IgG Fc receptors (FcgammaRs). Systemic lupus erythematosus is a prototype of IC-mediated autoimmune disease; thus, imbalance of these two types of FcgammaRs is probably involved in pathogenesis. However, how and to what extent each FcgammaR contributes to the disease remains unclear. In lupus-prone BXSB mice, while stimulatory FcgammaRs are intact, inhibitory FcgammaRIIB expression is impaired because of promoter region polymorphism. To dissect roles of stimulatory and inhibitory FcgammaRs, we established two gene-manipulated BXSB strains: one deficient in stimulatory FcgammaRs (BXSB.gamma(-/-)) and the other carrying wild-type Fcgr2b (BXSB.IIB(B6/B6)). The disease features were markedly suppressed in both mutant strains. Despite intact renal function, however, BXSB.gamma(-/-) had IC deposition in glomeruli associated with high-serum IgG anti-DNA Ab levels, in contrast to BXSB.IIB(B6/B6), which showed intact renal pathology and anti-DNA levels. Lymphocytes in BXSB.gamma(-/-) were activated, as in wild-type BXSB, but not in BXSB.IIB(B6/B6). Our results strongly suggest that both types of FcgammaRs in BXSB mice are differently involved in the process of disease progression, in which, while stimulatory FcgammaRs play roles in effecter phase of IC-mediated tissue inflammation, the BXSB-type impaired FcgammaRIIB promotes spontaneous activation of self-reactive lymphocytes and associated production of large amounts of autoantibodies and ICs. 相似文献
993.
Microbial production of 1,3-propanediol by Klebsiella pneumoniae using crude glycerol from biodiesel preparations 总被引:3,自引:0,他引:3
1,3-Propanediol (1,3-PD) was produced by Klebsiella pneumoniae using crude glycerol obtained from biodiesel production. The 1,3-PD concentration of 51.3 g/l−1 on crude glycerol from alkali-catalyzed methanolysis of soybean oil was comparable to that of 53 g/l−1 on crude glycerol derived from a lipase-catalyzed process. The productivities of 1.7 g l−1 h−1 on crude glycerol were comparable to that of 2 g l−1 h−1 on pure glycerol. It could be concluded that the crude glycerol could be directly converted to 1,3-PD without any prior purification. 相似文献
994.
探讨人核糖核酸抑制因子 (hRI)基因在人脐血干细胞中的转染及表达情况 ,及转染后对小鼠B16黑色素瘤生长的影响。用免疫磁珠分离系统 (MACS)分离纯化人脐血CD34+ 细胞后 ,用制备的含hRI基因的逆转录病毒上清转染脐血CD34+ 细胞 ,采用克隆形成法和PCR法检测转染效率 ,Western blot和免疫荧光法检测基因表达 ,同时观察RI对荷瘤C57BL小鼠B16黑色素瘤生长的影响。应用MACS能高度纯化人脐血CD34+ 细胞 ,使分选后的脐血CD34+ 细胞纯度平均达96.15%。hRI基因能够转染到脐血CD34+ 细胞上 ,转染效率达 35% ,Western blot和免疫荧光检测转染后CD34+ 细胞hRI基因有阳性表达。经转hRICD34+ 细胞治疗 ,使小鼠B16黑色素瘤的生长速度减慢 ,成瘤率和瘤重降低 ,成瘤潜伏期延长。 相似文献
995.
Resveratrol is a natural phenol with protective effects against cancer and inflammation-related diseases. Its mechanism of action involves the activation of nuclear factor E2 p45-related factor 2 (Nrf2), which plays a key role in regulation of genes driven by antioxidant response element (ARE). Inspired by the effect of resveratrol, here we synthesized a series of imine resveratrol analogs (IRAs), evaluated their abilities to activate Nrf2 by using cell based ARE-reporter assay. After the first-round screening, preliminary and quantitative structure-activity relationship (SAR) was analyzed, and the structural features determining Nrf2 activation ability were proposed. Two novel IRAs were designed and subsequently synthesized, namely 2-methoxyl-3,6-dihydroxyl-IRA and 2,3,6-trihydroxyl-IRA. They were proved to be the most potent Nrf2 activators among the IRAs. 相似文献
996.
997.
Liu XH Zhang ZY Andersson KB Husberg C Enger UH Ræder MG Christensen G Louch WE 《Cell calcium》2011,49(4):201-207
Reduced sarco(endo)plasmic reticulum (SR) Ca(2+) ATPase (SERCA2) contributes to the impaired cardiomyocyte Ca(2+) homeostasis observed in heart failure. We hypothesized that a reduction in SERCA2 also elicits myocardial ER/SR stress responses, including unfolded protein responses (UPR) and cardiomyocyte apoptosis, which may additionally contribute to the pathophysiology of this condition. Left ventricular myocardium from mice with cardiomyocyte-specific tamoxifen-inducible disruption of Serca2 (SERCA2 KO) was compared with aged-matched controls. In SERCA2 KO hearts, SERCA2 protein levels were markedly reduced to 2% of control values at 7 weeks following tamoxifen treatment. Serca2 disruption caused increased abundance of the ER stress-associated proteins CRT, GRP78, PERK, and eIF2α and increased phosphorylation of PERK and eIF2α, indicating UPR induction. Pro-apoptotic signaling was also activated in SERCA2 KO, as the abundance of CHOP, caspase 12, and Bax was increased. Indeed, TUNEL staining revealed an increased fraction of cardiomyocytes undergoing apoptosis in SERCA2 KO. ER-Tracker staining additionally revealed altered ER structure. These findings indicate that reduction in SERCA2 protein abundance is associated with marked ER/SR stress in cardiomyocytes, which induces UPR, apoptosis, and ER/SR structural alterations. This suggests that reduced SERCA2 abundance or function may contribute to the phenotype of heart failure also through induction of ER/SR stress responses. 相似文献
998.
999.
Xiaoming Song Jinpeng Wang Nan Li Jigao Yu Fanbo Meng Chendan Wei Chao Liu Wei Chen Fulei Nie Zhikang Zhang Ke Gong Xinyu Li Jingjing Hu Qihang Yang Yuxian Li Chunjin Li Shuyan Feng He Guo Jiaqing Yuan Qiaoying Pei Tong Yu Xi Kang Wei Zhao Tianyu Lei Pengchuan Sun Li Wang Weina Ge Di Guo Xueqian Duan Shaoqi Shen Chunlin Cui Ying Yu Yangqin Xie Jin Zhang Yue Hou Jianyu Wang Jinyu Wang Xiu‐Qing Li Andrew H. Paterson Xiyin Wang 《Plant biotechnology journal》2020,18(6):1444-1456
1000.
Yi Zhang Meng‐Chen Hou Jing‐Jing Li Ying Qi Yu Zhang Gang She Yu‐Jie Ren Wei Wu Zheng‐Da Pang Wenjun Xie Xiu‐Ling Deng Xiao‐Jun Du 《Journal of cellular and molecular medicine》2020,24(15):8505-8517
Activation of the sympatho‐β‐adrenergic receptors (β‐ARs) system is a hallmark of heart failure, leading to fibrosis and arrhythmias. Connexin 43 (Cx43) is the most abundant gap junctional protein in the myocardium. Current knowledge is limited regarding Cx43 remodelling in diverse cell types in the diseased myocardium and the underlying mechanism. We studied cell type‐dependent changes in Cx43 remodelling due to β‐AR overactivation and molecular mechanisms involved. Mouse models of isoproterenol stimulation or transgenic cardiomyocyte overexpression of β2‐AR were used, which exhibited cardiac fibrosis and up‐regulated total Cx43 abundance. In both models, whereas Cx43 expression in cardiomyocytes was reduced and more laterally distributed, fibroblasts exhibited elevated Cx43 expression and enhanced gap junction communication. Mechanistically, activation of β2‐AR in fibroblasts in vitro elevated Cx43 expression, which was abolished by the β2‐antagonist ICI‐118551 or protein kinase A inhibitor H‐89, but simulated by the adenylyl cyclase activator forskolin. Our in vitro and in vivo data showed that β‐AR activation‐induced production of IL‐18 sequentially stimulated Cx43 expression in fibroblasts in a paracrine fashion. In summary, our findings demonstrate a pivotal role of β‐AR in mediating distinct and cell type‐dependent changes in the expression and distribution of Cx43, leading to pathological gap junction remodelling in the myocardium. 相似文献