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61.
Genes of human mitochondrial tRNALeu(UUR) (mtRNALeu(UUR)) and its mutant (mtRNALeu(M)) were synthesized and inserted into the plasmid pGEM-9Zf(-) respectively.E.coli JM 109 was transformed by the recombinant plasmids containing the target genes. The mtRNALeu(UUR) and mtRNALeu(M) were expressed up to 19.10% and 17.76% of total small RNA respectively. They were purified to 54% homogeneity by DEAE-sepharose-CL4B column chromatography and finally repurified by 15% PAGE/urea. Their kinetic parameters forE.coli LeuRS were measured. The results showed that the value of kcal/ Km of mtRNALeu(M) was about one fifth of that of mtRNALeu(UUR) and indicated the leucine acceptability of mtRNALeu(M) was much lower than that of mtRNALeu(UUR).  相似文献   
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异源多角体蛋白对家蚕核型多角体病毒粒子的包装   总被引:1,自引:0,他引:1  
利用PCR方法从AcMNPV基因组DNA中分离出多角体蛋白基因 ,将该扩增片段克隆到转移载体pBacPAK8中 ,得到重组转移载体pOAc。将该质粒DNA与线性化的Bm BacPAK6病毒基因组DNA共传染BmN细胞 ,得到了能形成多角体且不产生蓝色空斑的重组病毒hp BmNPV。纯化该重组病毒的多角体颗粒 ,并对多角体蛋白、病毒核酸及多角体病毒颗粒进行分析 ,发现AcMNPV的多角体蛋白能在家蚕细胞中大量表达且能在细胞内识别家蚕核型多角体病毒并组装成多角体颗粒 ;病毒基因组DNA因部分交换 ,其酶切行为发生了相应的变化 ;电镜观察发现经AcMNPV多角体蛋白包装的家蚕核型多角体病毒的多角体颗粒大小为1 2 μm~ 2 9μm ,明显小于野生型家蚕核型多角体病毒的多角体颗粒  相似文献   
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Jia  Xin  Zha  Tianshan  Wang  Shan  Bourque  Charles P.-A.  Wang  Ben  Qin  Shugao  Zhang  Yuqing 《Plant and Soil》2018,429(1-2):437-450
Plant and Soil - Plant growth-promoting bacteria of the genus Bacillus are known to solubilize phosphates and enhance plant growth in many plant species. We explored the effects of the inoculation...  相似文献   
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Environmental health is an essential component of quality of life in modern societies. Foliar dust contains polycyclic aromatic hydrocarbons (PAHs) that may have harmful effects on human health. The PAHs concentration of foliar dust is useful to assess environmental air pollution. Our results indicate that: (1) the highest levels of PAHs were distributed in urban areas, with a mean of 3430.23 ng·g?1, lower mean concentrations were found in suburban (2282.12 ng·g?1), and rural areas (1671.06 ng·g?1). (2) Diagnostic ratios and principal component analysis were used to identify the sources of PAHs: Gasoline vehicle traffic emissions were the predominant source in urban areas, along with coal and coke combustion. In suburban areas, the main sources were petroleum combustion (especially liquid fossil fuels) and coal combustion. Coal and wood combustion were the primary source of PAHs in foliar dust in rural areas. (3) The incremental lifetime cancer risk (ILCR), estimated based on the results of this study indicate that urban residents were potentially exposed to high cancer risk via both dust ingestion and dermal contact. We conclude that urbanization has significant effects on the PAH concentrations of foliar dust, illustrating the importance of trees in improving air quality in urban areas.  相似文献   
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Vγ9Vδ2 T cells are a minor subset of lymphocytes in the peripheral blood that has been extensively investigated for their tolerability, safety and anticancer efficacy. A hindrance to the broad application of these cells for adoptive cellular immunotherapy has been attaining clinically appropriate numbers of Vγ9Vδ2 T cells. Furthermore, Vγ9Vδ2 T cells exist at low frequencies among cancer patients. We, therefore, sought to conceive an economical method that allows for a quick and robust large-scale expansion of Vγ9Vδ2 T cells. A two-step protocol was developed, in which peripheral blood mononuclear cells (PBMCs) from healthy donors or cancer patients were activated with Zometa and interleukin (IL)-2, followed by co-culturing with gamma-irradiated, CD64-, CD86- and CD137L-expressing K562 artificial antigen-presenting cells (aAPCs) in the presence of the anti-CD3 antibody OKT3. We optimized the co-culture ratio of K562 aAPCs to immune cells, and migrated this method to a G-Rex cell growth platform to derive clinically relevant cell numbers in a Good Manufacturing Practice (GMP)-compliant manner. We further include a depletion step to selectively remove αβ T lymphocytes. The method exhibited high expansion folds and a specific enrichment of Vγ9Vδ2 T cells. Expanded Vγ9Vδ2 T cells displayed an effector memory phenotype with a concomitant down-regulated expression of inhibitory immune checkpoint receptors. Finally, we ascertained the cytotoxic activity of these expanded cells by using nonmodified and chimeric antigen receptor (CAR)–engrafted Vγ9Vδ2 T cells against a panel of solid tumor cells. Overall, we report an efficient approach to generate highly functional Vγ9Vδ2 T cells in massive numbers suitable for clinical application in an allogeneic setting.  相似文献   
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