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991.
Shiming Lin Chung-Hung Hong Bor-Ching Sheu Long-Xin Wu Wan-Chen Huang Wei-Chih Huang Cheng-Yan Guo 《Biochemistry and Biophysics Reports》2016
This paper proposes an effective approach to distinguish whether samples include Human Papilloma virus type-16 (HPV16) by Atomic force microscopy (AFM). AFM is an important instrument in nanobiotechnology field. At first we identified the HPV16 by Polymerase chain reaction (PCR) analysis and Western blotting from specimen of the HPV patient (E12) and the normal (C2), and then we used an AFM to observe the surface ultrastructure by tapping mode and to measure the unbinding force between HPV16 coupled to an AFM tip and anti-HPV16 L1 coated on the substrate surface by contact mode. The experimental results by tapping mode show that the size of a single HPV viron was similar to its SEM image from the previous literatures; moreover, based on the purposed methods and the analysis, two obvious findings that we can determine whether or not the subject is a HPV patient can be derived from the results; one is based on the distribution of unbinding forces, and the other is based on the distribution of the stiffness. Furthermore, the proposed method could be a useful technique for further investigating the potential role among subtypes of HPVs in the oncogenesis of human cervical cancer. 相似文献
992.
Genetic mutation analysis at early stages of cell line development using next generation sequencing
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Chapman Wright Joost Groot Samantha Swahn Helen McLaughlin Mei Liu Chongfeng Xu Chao Sun Eric Zheng Scott Estes 《Biotechnology progress》2016,32(3):813-817
A central goal for most biopharmaceutical companies is to reduce the development timeline to reach clinical proof of concept. This objective requires the development of tools that ensure the quality of biotherapeutic material destined for the clinic. Recent advances in high throughput protein analytics provide confidence in our ability to assess productivity and product quality attributes at early stages of cell line development. However, one quality attribute has, until recently, been absent from the standard battery of analytical tests facilitating informed choices early in cell line selection: genetic sequence confirmation. Techniques historically used for mutation analysis, such as detailed mass spectrometry, have limitations on the sample number and turnaround times making it less attractive at early stages. Thus, we explored the utility of Next‐Generation Sequencing (NGS) as a solution to address these limitations. Amplicon sequencing is one such NGS technique that is robust, rapid, sensitive, and amenable to multiplexing, all of which are essential attributes for our purposes. Here we report a NGS method based upon amplicon sequencing that has been successfully incorporated into our cell line development workflow alongside other high‐throughput protein analytical assays. The NGS method has demonstrated its value by identifying at least one Chinese hamster ovary (CHO) clone expressing a variant form of the biotherapeutic in each of the four clinical programs in which it has been utilized. We believe this sequence confirmation method is essential to safely accelerating the time to clinical proof of concept of biotherapeutics, and guard against delays related to sequence mutations. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:813–817, 2016 相似文献
993.
从阿维菌素获得诺贝尔奖到中国创造 总被引:1,自引:0,他引:1
纵观世界历史,大国崛起无不伴随着科技的兴起和机制体制的突破。来自大自然的天然产物对于人类的健康起到非常重要的作用,从抗肿瘤明星分子紫杉醇到挽救了无数人生命的抗感染药物青霉素,从治疗代谢疾病到营养保健,都离不开天然产物。此外,还有大量天然产物资源没有被开发过。而随着阿维菌素的发现者Satoshiōmura教授和William C.Campbell博士,及青蒿素的发现者屠呦呦研究员因为这两种天然产物在治疗寄生虫感染病和疟疾上的应用而获得2015年诺贝尔生理与医学奖后,天然产物有望迎来其发展的第二个黄金时代。我国是世界工厂也是天然药物的资源大国,为了实现产业升级,弯道超车,实现大国崛起的中国梦,我国科学家在"十二五"期间围绕着天然产物的高产和创新两大主题开展了富有成效的合成生物学研究。阿维菌素是由阿维链霉菌产生的高效低毒生物杀虫剂,其原料产能占国际市场的100%。但我国原有生产菌株的单位发酵产量较低,高消耗、高污染、片面追求生产规模的粗放型发展模式已经成了阻碍低碳经济持续高速发展的瓶颈。如何从根本上解决问题,提高生产菌株的单位发酵产量和原料利用率,降低能耗和生产成本,减少环境污染,是促进我国从"发酵大国"向"发酵强国"转变的关键。本文以阿维菌素为例,综述其基础研究的技术发展,特别是中国科学院微生物研究所引入合成生物学技术,将阿维菌素的单位产量提高了1000倍,至9 g/L,在内蒙古新威远与阿维菌素产业联盟的公司应用,迫使默克公司全面退出阿维菌素历史舞台,从而引领产业迅速发展的过程,为我国其它天然产物生物制造品种的改良提供思路和方法。 相似文献
994.
Priyanka Prakash Yong Zhou Hong Liang John?F. Hancock Alemayehu?A. Gorfe 《Biophysical journal》2016,110(5):1125-1138
K-Ras is a membrane-associated GTPase that cycles between active and inactive conformational states to regulate a variety of cell signaling pathways. Somatic mutations in K-Ras are linked to 15–20% of all human tumors. K-Ras attaches to the inner leaflet of the plasma membrane via a farnesylated polybasic domain; however, the structural details of the complex remain poorly understood. Based on extensive (7.5 μs total) atomistic molecular dynamics simulations here we show that oncogenic mutant K-Ras interacts with a negatively charged lipid bilayer membrane in multiple orientations. Of these, two highly populated orientations account for ∼54% of the conformers whose catalytic domain directly interacts with the bilayer. In one of these orientation states, membrane binding involves helices 3 and 4 of the catalytic domain in addition to the farnesyl and polybasic motifs. In the other orientation, β-strands 1–3 and helix 2 on the opposite face of the catalytic domain contribute to membrane binding. Flexibility of the linker region was found to be important for the reorientation. The biological significance of these observations was evaluated by initial experiments in cells overexpressing mutant K-Ras as well as by an analysis of Ras-effector complex structures. The results suggest that only one of the two major orientation states is capable of effector binding. We propose that the different modes of membrane binding may be exploited in structure-based drug design efforts for cancer therapy. 相似文献
995.
Figen Seymen Youn Jung Kim Ye Ji Lee Jenny Kang Tak-Heun Kim Hwajung Choi Mine Koruyucu Yelda Kasimoglu Elif Bahar Tuna Koray Gencay Teo Jeon Shin Hong-Keun Hyun Young-Jae Kim Sang-Hoon Lee Zang Hee Lee Hong Zhang Jan C-C. Hu James P. Simmer Eui-Sic Cho Jung-Wook Kim 《American journal of human genetics》2016,99(5):1199-1205
996.
Taotao Feng Xiaoyu ChenXiuwen Qiao Zhao SunHaining Wang Yu Qi Chenglin Hong 《Analytical biochemistry》2016
In this work, a simple electrochemical immunosensor was developed for the detection of carcinoembryonic antigen (CEA) based on rhombic dodecahedral Cu2O nanocrystals–graphene oxide–gold nanoparticles (rCu2O–GO–AuNPs). GO as the template and surfactant resulting in rCu2O exhibit improved rhombic dodecahedral structure uniformity and excellent electrochemical performance. Moreover, GO was found to be able to effectively improve the long stability of rCu2O on the electrode response. Under optimal conditions, the immunosensor showed a low limit of detection (0.004 ng ml−1) and a large linear range (0.01–120 ng ml−1). This work presents a potential alternative for the diagnostic applications of GO-supported special morphology materials in biomedicine and biosensors. 相似文献
997.
Isothermal and rapid amplification detection of nucleic acids is an important technology in environmental monitoring, foodborne pathogen detection, and point-of-care clinical diagnostics. Here we have developed a novel method of isothermal signal amplification for single-stranded DNA (ssDNA) detection. The ssDNA target could be used as an initiator, coupled with a double-nicked molecular beacon, to originate amplification cycles, achieving cascade signal amplification. In addition, the method showed good specificity and strong anti-jamming capability. Overall, it is a one-pot and isothermal strand displacement amplification method without the requirement of a stepwise procedure, which greatly simplifies the experimental procedure and decreases the probability of contamination of samples. With its advantages, the method would be very useful to detect nucleic acids in point-of-care or field use. 相似文献
998.
Ancillary contributions of heterologous biotin protein ligase and carbonic anhydrase for CO2 incorporation into 3‐hydroxypropionate by metabolically engineered Pyrococcus furiosus
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999.
1000.
Genomic constitution and taxonomy of the Chinese hexaploids Elymus cylindricus and E. breviaristatus (Poaceae: Triticeae)
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Cai‐Rong Yang Bernard‐R. Baum Wei‐Huan Chen Hai‐Qin Zhang Xiao‐Yan Liu Xing Fan Li‐Na Sha Hou‐Yang Kang Yi Wang Yong‐Hong Zhou 《Botanical journal of the Linnean Society. Linnean Society of London》2016,182(3):650-657
Elymus cylindricus (2n = 6x = 42) and E. breviaristatus (2n = 6x = 42) are distributed in grasslands and deserts of northern and north‐western China. Their genomic constitution and taxonomic status are unclear. Elymus cylindricus was crossed with E. wawawaiensis J.R.Carlson & Barkworth ( StH ), Roegneria grandis Keng ( StY ) and Campeiostachys dahurica (Turcz. ex Griseb.) B.R.Baum, J.L. Y ang & C. Y en var. dahurica ( StYH ). Meiotic pairing in the hybrids E. cylindricus × E. wawawaiensis ( StH ), E. cylindricus × R. grandis ( StY ) and E. cylindricus × C. dahurica var. dahurica ( StYH ) showed on average 10.00, 11.30 and 20.92 bivalents per cell, respectively. Elymus breviaristatus was crossed with C. dahurica var. dahurica ( StYH ) and E. cylindricus. Chromosome pairing in the hybrids of E. breviaristatus × C. dahurica var. dahurica and E. breviaristatus × E. cylindricus showed on average 19.60 and 19.27 bivalents, respectively. Genomic in situ hybridization (GI SH ) revealed the presence of St , Y and H genomes in E. cylindricus and E. breviaristatus. An intergenomic rearrangement was observed in E. cylindricus using GI SH . Meiotic pairing data and GI SH indicated that both E. cylindricus and E. breviaristatus are allohexaploids containing the StYH genomes. Elymus cylindricus and E. breviaristatus should be treated as Campeiostachys dahurica var. cylindrica and Campeiostachys breviaristata, respectively. 相似文献