生物膜研究在生物物理研究所的兴起与发展

生物膜研究是现代生物学研究的前沿方向之一,本文对文革结束以后生物膜研究在中国科学院生物物理研究所(下称生物物理所)的兴起与发展进行了系统回顾.文革结束后,中国科学院领导了解到国外生物膜研究迅速发展的情况,迅即派遣以生物物理所杨福愉为组长,包括生物化学研究所、动物研究所、植物研究所、上海实验生物研究所等六人的代表团前往联邦德国进行考察.考察结束后院领导根据多学科交叉对生物膜研究的重要作用,组织了生物物理所、植物所、中国医学科学院、北京医科大学等单位联合申报国家自然科学基金委员会重大项目"膜脂-膜蛋白的相互作用及其在医学和农业上的应用",并获得立项.与此同时,院领导建议由中国生物物理学会、中国生化学会和中国细胞生物学会共同组织生物膜学术讨论会.首次会议于1979年3月在北京友谊宾馆举行,以后每3年召开1次,从未中断,有力地促进了生物膜研究的交流与发展.2003年举行的第200届香山会议专门组织讨论21世纪生物膜研究在中国的布局,进一步推动了生物膜研究的发展.本文还重点阐述了中国科学院生物物理研究所在生物膜研究方面所取得的代表性成果:a.金属离子通过膜脂-膜蛋白相互作用调控生物膜能量转换、物质运输及信号转导的分子机制;b.提出"克山病是一种心肌线粒体病"的重要观点;c.发现溶酶体内含有为量甚微、一般认为是消化酶的胰凝乳蛋白酶,并阐明了它通过溶酶体膜外泄后参与细胞凋亡的作用机制;d.确定了通过调控线粒体动态变化而干预肿瘤细胞迁移侵袭的新靶标.最后,特别值得一提的是,2004年,常文瑞与植物研究所匡廷云等在《自然》(Nature)发表了《菠菜中主要捕光色素复合体2.72魡分辨率的晶体结构分析》的研究论文,2005年饶子和与徐建兴等在《细胞》(Cell)发表了《猪心线粒体呼吸链复合体Ⅱ的晶体结构》的研究论文,充分标志着我国生物膜研究已在国际上占据一席之地.2004年,徐涛研究员因其在囊泡转运方面的丰厚学术积淀,作为首席科学家组织一批生物膜专家承担了国内首个生物膜973项目,这标志着国内生物膜研究开始进入一个新的发展时期.     

碱基编辑技术的发展与应用

碱基编辑技术,以CRISPR/Cas系统为平台,引导胞嘧啶脱氨酶或腺嘌呤脱氨酶至特定的基因组靶点,产生靶向性的C至T或者A至G的碱基转换。自碱基编辑技术问世以来,全球多个科研团队通过优化改进得到了一系列高精准性、广靶向性、小编辑框、普适性的碱基编辑器。在应用方面,碱基编辑器能够在人体细胞、动植物细胞以及胚胎中进行高效的碱基转换,在治疗人类遗传病、构建动物疾病模型、植物育种等方面具有巨大的应用潜能。本文就碱基编辑技术的发展、优化和应用等方面进行综述和展望。     

“人造精子”介导基因编辑技术的建立与应用

"人造精子",即小鼠孤雄单倍体胚胎干细胞,是一种可在体外长期培养扩增和进行遗传操作的细胞,因其具备胚胎干细胞自我更新和多向分化的特性,并拥有代替精子使卵母细胞"受精",产生半克隆小鼠的能力,而被广泛应用于生命科学研究的各个领域。与CRISPR-Cas9技术相结合,"人造精子"能够同时实现细胞水平和动物水平的复杂基因编辑,因此可作为一个有力的研究工具。本文就"人造精子"技术的建立与改进、在基因编辑方面的应用,以及全基因组标签计划的提出与进展展开综述。     

低氧暴露所致大鼠骨骼肌萎缩的蛋白转化调节机制

目的 对比低氧暴露和常氧下配对低氧摄食干预(半饥饿状态)下大鼠骨骼肌蛋白质合成和分解相关基因表达的差异,以探讨低氧暴露诱导骨骼肌萎缩发生的可能机制。方法 SD大鼠分为:①常氧正常饮食组(C组);②低氧正常饮食组(H组),氧气浓度为12.4%;③常氧配对饮食组(P组),投食量即为H组前一天摄食量。4周干预后测量大鼠体成分,取比目鱼肌(SOL)和趾长伸肌(EDL),称量湿重;HE染色观察肌纤维形态,计算肌纤维横截面积(FCSA);WB测试骨骼肌中HIF1α、Akt、p-Akt及骨骼肌蛋白合成和分解相关基因蛋白含量。结果 1)H组大鼠体重较C组持续下降,P组与C组间无显著性差异;干预初期H组(P组同)摄食量较C组显著下降,后期两组间无差异;(2)干预后,H组大鼠体质量和肌肉总量较C组和P组显著性降低,P组与C组间无差异;H组两肌肉湿重较C组显著下降;H组EDL的FCSA显著低于C组和P组;(3)H组EDL中HIF1α蛋白含量显著高于C组;H组和P组SOL中p-Akt/Akt比值显著低于C组;H组EDL中mTOR、4EBP1蛋白含量显著低于C组,atrogin1、MuRF1、beclin1蛋白含量及LC3Ⅱ/Ⅰ比值显著高于C组,H组SOL中MuRF1蛋白含量显著高于C组和P组。结论 低氧所致的骨骼肌萎缩由低氧特异性因素诱发,表现为以快肌为主的骨骼肌蛋白合成减少和分解增加,而非低氧下摄食量减少引起。     

一种强力霉素诱导型白念珠菌基因敲除与筛选标记再循环工具包（英文）

【目的】在白念珠菌中建立一个快捷方便经济的基因敲除与筛选标记再循环的DNA操作系统。【方法】通过ExoIII介导的不依赖于连接酶的克隆策略,在异源筛选标记基因CmLEU2、CdHIS1和CdARG4基因的两侧分别插入了loxP位点,成为筛选标记基因盒扩增的模板。全基因合成了经过白念珠菌密码子优化的rTetR元件,并组装成Tet-on启动子。将密码子优化的重组酶Cre基因置于该启动子控制下。然后将他们插入筛选标记基因CdHIS1和CdARG4的CDS区域,形成筛选标记基因再循环载体。【结果】构建了3个用于白念珠菌基因敲除的侧翼含有loxP位点的筛选标记基因载体,以及2个含有Tet-on启动子控制的Cre酶的载体用于筛选标记基因的再循环。【结论】成功构建了一个白念珠菌中可诱导的基因敲除和筛选标记再循环的载体系统并成功应用于多个基因缺失株构建。这个系统有助于快速构建白念珠菌的单基因和多基因敲除菌株。     

Structure and specific DNA binding of the rat liver glucocorticoid receptor

During recent years major advances have been made in our understanding of glucocorticoid mechanism of action. This progress has been made possible by access to purified glucocorticoid receptor in significant amounts as well as by application of hybrid DNA technology within the field of glucocorticoid control of gene expression. Especially the mammary tumour virus genome has turned out to be a convenient experimental system suitable for such investigations. This paper summarizes some of the work carried out in our own laboratory, partially in collaboration with Dr Keith Yamamoto and his associates at the Department of Biochemistry and Biophysics, University of California, San Francisco, U.S.A.     

The 2009 Nobel Prize in Chemistry: Thomas A. Steitz and the structure of the ribosome

Over the past 200 years, there have been countless groundbreaking discoveries in biology and medicine at Yale University. However, one particularly noteworthy discovery with profoundly important and broad consequences happened here in just the past two decades. In 2009, Thomas Steitz, the Sterling Professor of Molecular Biophysics & Biochemistry, was awarded the Nobel Prize in Chemistry for "studies of the structure and function of the ribosome," along with Venkatraman Ramakrishnan of the MRC Laboratory of Molecular Biology and Ada E. Yonath of the Weizmann Institute of Science. This article covers the historical context of Steitz's important discovery, the techniques his laboratory used to study the ribosome, and the impact that this research has had, and will have, on the future of biological and medical research.     

孕烷X受体配体结合结构域蛋白晶体的X线衍射结构解析

目的:利用X线衍射技术解析孕烷X受体(PXR)配体结合结构域(LBD)蛋白晶体的3维结构。方法:对PXR蛋白LBD(130~434氨基酸残基)序列进行密码子优化并化学合成后克隆至pRSFDuet-1表达载体,再将载体导入大肠杆菌BL21(DE3),对PXR-LBD蛋白进行原核表达与分离纯化;采用晶体筛选试剂盒筛选蛋白结晶条件,采用悬滴法获得目标蛋白的晶体;对获得的蛋白晶体进行X线晶体衍射检测,并收集相关数据建立PXR-LBD的三维结构。结果:获得了PXR-LBD的高质量晶体并利用X线衍射解析了该蛋白质晶体的结构数据,使用Phenix.refine软件和COOT软件等对结构进行修正,最终获得了高分辨率的3维结构数据。结论:完成了孕烷X受体配体结合结构域蛋白晶体的X线衍射结构解析,为研究和开发PXR相关药物奠定了基础。     

The effect of krypton ions on Artemia cysts

Summary Dry eggs of Artemia salina were exposed to 80 KV-X-rays and Kr-Ions (energy: 11.75 MeV/u; LET: 3500 KeV/µm). Emergence, hatching and survival were determined. The results demonstrate a very high resistance both to sparsely and ionizing radiation.On leave from Institute of Biophysics, Academia Sinica, Beijing, People's Republic of China     

The Use of the Antigenic Protein HPV6 L1 to Induce the Synthesis of Interferon,CD4 and CD8 T Lymphocytes,and Granzyme B in Blood and Splenocytes of Mice in Order to Develop a Broad Vaccine against Anogenital Papillomatoses

Doklady Biochemistry and Biophysics - The antigenic protein HPV6 L1 was synthesized in the plant expression system based on transgenic tomato fruit during the development of an oral vaccine against...     

Jordan Raff

Jordan Raff is a Cancer Research UK funded group leader at the Wellcome/CR-UK Gurdon Institute in Cambridge, England. He obtained his PhD from the Department of Biochemistry at Imperial College, London, and he worked as a Post-doctoral fellow at the Department of Biophysics and Biochemistry, University of California, San Francisco. He is currently a Director of the Company of Biologists, and on the committee of the British Society of Cell Biology. He has studied centrosomes and cell division in fruit flies throughout his scientific career.     

The Synthesis of Main Capsid Protein of Anogenital Type HPV6 L1 in Plant Expression System on the Basis of Tomato Fruits

Doklady Biochemistry and Biophysics - The anogenital type HPV6 L1 major capsid protein was synthesized in a plant expression system on the basis of tomato fruits. The content of HPV6 L1 reached 380...     

Bacterial rhodopsins: evolution of a mechanistic model for the ion pumps

Walther Stoeckenius received a MD degree at the University of Hamburg, Germany in 1950. After 18 months of clinical work as an intern, he began postdoctoral work on the development of pox viruses at the Institute for Tropical Medicine in Hamburg using mainly electron microscopy techniques. After two years he moved as Assistant Professor to the Department of Pathology at the University of Hamburg and became Docent for Pathology in 1958. In addition to teaching and routine pathology work, he continued to use electron microscopy to explore the fine structure of cells and developed an interpretation of the triple-layered appearance of membranes in electron micrographs in terms of molecular structure and the chemistry of osmium tetroxide fixation. In 1959 he obtained a position as Research Associate in Keith Porter's laboratory at Rockefeller University. This was changed after a few months to Assistant Professor and he stayed there, later as Associate Professor, for eight years. The work on membrane structure continued, and a model was developed that described the membrane as a lipid bilayer with embedded protein domains. In efforts to isolate such domains, the purple membrane and bacteriorhodopsin were discovered. In 1966, the lure of California became irresistible and Dr. Stoeckenius accepted a professorship at the University of California at San Francisco. The work on bacteriorhodopsin continued there with the emphasis changing from electron microscopy to spectroscopy and biochemical techniques. He is now Professor Emeritus there in the Department of Biochemistry and Biophysics and the Cardiovascular Research Institute.     

Temperature Dependence of Tryptophan Fluorescence Lifetime as an Indicator of Its Microenvironment Dynamics

Doklady Biochemistry and Biophysics - The spectral-kinetic characteristics of the fluorescence of the tryptophan molecule in an aqueous solution and in the composition of a protein (albumin) were...     

Oxidative Mechanisms in Neurodegenerative Disorders. Proceedings from a satellite symposium, Guilin, China, 7-11 February 2004

This special issue of Molecular Neurobiology presents a series of mini-reviews resulting from an ISN satellite symposium entitled Oxidative Mechanisms in Neurodegenerative Disorders Held at the Guilin Park Hotel, Guilin, China on February 7-11, 2004. The timely topics in this symposium were contributed by more than 20 internationally acclaimed scientists and attended by more than 100 participants interested in this subject matter. This satellite symposium was also part of the 6th Biennial Asian-Pacific Society for Neurochemistry (APSN) meeting held in Hong Kong, China. Organizers included Dr. Piu Chan (Capital University of Medical Sciences and Beijing Institute of Geriatrics) Dr. Albert Y. Sun and Dr. Grace Y. Sun (University of Missouri, Columbia, MO). The satellite meeting was hosted by Xuanwu Hospital of Capital University of Medical Sciences and Guilin Medical College, and was generously supported by the following: the International Society of Neurochemistry (ISN), National Institutes of Health (USA) (1 R13 NS047414), the Geriatric Research, Education and Clinical Center (GRECC) VA Medical Center, Minneapolis, MN, USA, The Chinese Society of Neuroscience, Chinese Medical Association, Chinese Journal of Neurology, Shanghai Roche Pharmaceuticals Ltd., and Beijing QuiXave United Technology Ltd. Corporation.     

Among Antibody-Like Molecules,Monobodies, Able to Interact with Nucleocapsid Protein of SARS-CoV Virus,There Are Monobodies with High Affinity to Nucleocapsid Protein of SARS-CoV-2 Virus

Doklady Biochemistry and Biophysics - Seven amino acid sequences of antibody mimetics molecules, monobodies, capable of interacting with the nucleocapsid protein of the SARS-CoV virus, were taken...     

The Relationship between p53-Positive Neurons and Dark Neurons in the Hippocampus of Rats after Surgical Interventions on the Nasal Septum

Doklady Biochemistry and Biophysics - The study evaluates the dependence of p53 protein expression on the appearance of dark neurons (DNs) in the hippocampus in rats during experimental modeling of...     

TTK Isoforms Interact with Two Regions of the Mep-1 Protein of Drosophila melanogaster

Doklady Biochemistry and Biophysics - The Drosophila TTK protein is involved in the processes of cell differentiation and is represented by two isoforms, TTK69 and TTK88, which have a common...     

Antibodies to the N-Terminal Domain of Angiotensin-Converting Enzyme (ACE2) That Block Its Interaction with SARS-CoV-2 S Protein

Doklady Biochemistry and Biophysics - SARS-CoV-2 is a new coronavirus that is the cause of COVID-19 pandemic. To enter the cell, the virus interacts via its surface S protein with...