原子力显微镜对玉米黑粉菌及蛋白分子的研究
|
摘要
对细胞的表面形貌结构的认识是进一步了解细胞功能并探讨生命现象的重要基础。不同的研究技术和工具对研究起着重要的作用,其中原子力显微镜(AFM)有高分辨率和直观可视化的特点,它的样品制备简单,扫描时不会使生物样品发生较大的改变,还可以在近生理条件下观察生物细胞的形貌结构信息。这些特点使它迅速的应用于生物领域。
本论文一方面应用AFM和光学显微镜对玉米黑粉菌细胞的表面形貌结构进行了表征,并且观察了抑菌剂萎锈灵及潮霉素对黑粉菌细胞的表面形貌结构的影响,进一步的分析解释了药物对细胞的影响机理。另一方面通过AFM单分子力谱对芽孢衣蛋白相互作用进行了研究。论文内容主要体现在以下三个方面：
1.利用AFM对玉米黑粉菌单倍体时期生长的形态变化进行了观测,同时对空气中长期放置的黑粉菌进行了观察。通过AFM图可以清楚地观察到黑粉菌单倍体结合管生长的形态的变化及几十纳米级的菌体表面形貌。
2.通过AFM研究抑菌剂萎锈灵和潮霉素对玉米黑粉菌细胞形貌的影响。结果表明萎锈灵对黑粉菌不仅有生长的抑制作用,在黑粉菌菌体的表面随机出现了很多小坑。而潮霉素却使黑粉菌的表面变的比较平滑,粗糙度降低,没有出现坑。
3.利用单分子力谱对芽孢衣蛋白CotE间的相互作用进行了动力学研究。研究得出CotE蛋白分子间有弱的相互作用,在加载速率为3160 pNs-1时,其相互作用力的约为49.4pN,解离速率常数为0.146s-1,获得了其动力学信息。
Comprehension of the cell surface topography structure is important foundation to further understand the cell function and biological phenomena. Different tools and technology play an important role in researches. Atomic force microscope (AFM) is suitable for high-resolution imaging. Simple preparation for samples makes fewer changes on biological samples and the morphological structure of biological cells could be observed in physiological conditions.
The surface topography structure of Ustilago maydis haploid cells were characterized by AFM and optical microscopy. The effects of antibacterial agent hygromycin and Carboxin on topography structure of Ustilago maydis cells were observed. The interactions among proteins were also investigated by single molecule force spectroscopy. The content of article mainly includes the following three aspects.
1. Morpholoical changes of Ustilago maydis haploid and Ustilago maydis in long-term air exposure be observed via AFM. Morphological changes of combination tube in haploid and its dozen-nanometer-level morphology could be observed in AFM.
2. The effects of antibacterial agent hygromycin and Carboxin on morphology of Ustilago maydis were investigated by AFM. The results showed that Carboxin could not only inhibit the growth of Ustilago maydis, but also induced pits in cell surface. It was found that cell surface of Ustilago maydis become smoother, less rough, and no pits appeared after adding hygromycin.
3. The interactions between spore coat proteins were studied by single molecule force spectroscopy. The interaction force between CotE is 49.4 pN at loading rate of 3160 pNs-1. The dissociation rate constant is about 0.146s-1. The dynamic information was obtained.
本论文一方面应用AFM和光学显微镜对玉米黑粉菌细胞的表面形貌结构进行了表征,并且观察了抑菌剂萎锈灵及潮霉素对黑粉菌细胞的表面形貌结构的影响,进一步的分析解释了药物对细胞的影响机理。另一方面通过AFM单分子力谱对芽孢衣蛋白相互作用进行了研究。论文内容主要体现在以下三个方面：
1.利用AFM对玉米黑粉菌单倍体时期生长的形态变化进行了观测,同时对空气中长期放置的黑粉菌进行了观察。通过AFM图可以清楚地观察到黑粉菌单倍体结合管生长的形态的变化及几十纳米级的菌体表面形貌。
2.通过AFM研究抑菌剂萎锈灵和潮霉素对玉米黑粉菌细胞形貌的影响。结果表明萎锈灵对黑粉菌不仅有生长的抑制作用,在黑粉菌菌体的表面随机出现了很多小坑。而潮霉素却使黑粉菌的表面变的比较平滑,粗糙度降低,没有出现坑。
3.利用单分子力谱对芽孢衣蛋白CotE间的相互作用进行了动力学研究。研究得出CotE蛋白分子间有弱的相互作用,在加载速率为3160 pNs-1时,其相互作用力的约为49.4pN,解离速率常数为0.146s-1,获得了其动力学信息。
Comprehension of the cell surface topography structure is important foundation to further understand the cell function and biological phenomena. Different tools and technology play an important role in researches. Atomic force microscope (AFM) is suitable for high-resolution imaging. Simple preparation for samples makes fewer changes on biological samples and the morphological structure of biological cells could be observed in physiological conditions.
The surface topography structure of Ustilago maydis haploid cells were characterized by AFM and optical microscopy. The effects of antibacterial agent hygromycin and Carboxin on topography structure of Ustilago maydis cells were observed. The interactions among proteins were also investigated by single molecule force spectroscopy. The content of article mainly includes the following three aspects.
1. Morpholoical changes of Ustilago maydis haploid and Ustilago maydis in long-term air exposure be observed via AFM. Morphological changes of combination tube in haploid and its dozen-nanometer-level morphology could be observed in AFM.
2. The effects of antibacterial agent hygromycin and Carboxin on morphology of Ustilago maydis were investigated by AFM. The results showed that Carboxin could not only inhibit the growth of Ustilago maydis, but also induced pits in cell surface. It was found that cell surface of Ustilago maydis become smoother, less rough, and no pits appeared after adding hygromycin.
3. The interactions between spore coat proteins were studied by single molecule force spectroscopy. The interaction force between CotE is 49.4 pN at loading rate of 3160 pNs-1. The dissociation rate constant is about 0.146s-1. The dynamic information was obtained.
引文
[1]Service R F. Brightness speeds search for structures great and small [J]. Science,1997, 277(5330):1217-1219.
[2]Rost B. Marrying structure and genomics [J]. Structure,1998,6(3):259-263.
[3]Shapiro L, Lima C D. The argonne structural genomics workshop:lamaze class for the birth of a new science [J]. Structure,1998,6(3):265-267.
[4]Venter J C, Adams M D, Myers E W, et al. The sequence of the human genome [J]. Science, 2001,291(5507):1304-1351.
[5]Baltimore D. Our genome unveiled [J]. Nature,2001,409(6822):814-816.
[6]潘宪明.结构生物学——21世纪生命科学的重要前沿[J].中国科学基金,1998,(3)：157-160.
[7]邹承鲁.结构生物学的时代已经开始[J].科技导报,1995,(4)：7-11.
[8]Binnig G, Quate C F, Gerber C. Atomic force microscope [J]. Physical Review Letters, 1986,56(9):930-933.
[9]白春礼.扫描隧道显微术及其应用[M].1992.
[10]Weisenhorn a L, Hansma P K, Albrecht T R, et al. Forces in atomic force microscope in air and water [J]. Applied Physics Letters,1989,54(26):2651-2653.
[11]谭琰.原子力显微镜对生物大分子的研究[D].湘潭大学,2006.
[12]牟旭东.新型扫描力显微镜的研究[D].1998.
[13]陈伟.轻敲模式原子力显微镜的优化研究[D].浙江大学,2002.
[14]Binnig G, Rohrer H, Gerber C, et al. Surface studies by scanning tunneling microscopy [J]. Physical Review Letters,1982,49(1):57.
[15]程舒雯.扫描隧道显微镜性能优化及实用化研究[D].浙江大学,2003.
[16]马尚行.扫描隧道显微镜的新方法研究和系统研制[D].浙江大学,2005.
[17]Allison D P, Hinterdorfer P, Han W. Biomolecular force measurements and the atomic force microscope [J]. Current Opinion in Biotechnology,2002,13(1):47-51.
[18]张德添,何昆,张飒,et al.原子力显微镜发展近况及其应用[J].现代仪器,2002,(3)：6-9.
[19]白春礼.扫描力显微术[M].北京；科学出版社.2000：8-9.
[20]施春陵,蒋建清.原子力显微镜(AFM)在材料性能分析中的应用[J].江苏冶金,2005,(1)：7-9.
[21]鲍俊杰,高明志,周海峰, et al.原子力显微镜在聚氨酯材料性能分析中的应用[J].弹性体,2006,(3)：53-57.
[22]李琦,郑琪,丁焰,et al.原子力显微镜在染色体研究中的应用[J].遗传,2009,(6)：573-580.
[23]程敏熙.STM的同胞兄弟--原子力显微镜(AFM) [J]大学物理,2000,19(9)：5.
[24]Santos N C, Castanho M. An overview of the biophysical applications of atomic force microscopy [J]. Biophysical Chemistry,2004,107(2):133-149.
[25]Maurice P, Forsythe J, Hersman L, et al. Application of atomic-force microscopy to studies of microbial interactions with hydrous Fe(Ⅲ)-oxides [J]. Chemical Geology,1996, 132(1-4):33-43.
[26]Cabral J T, Higgins J S, Mcleish T C B, et al. Bulk spinodal decomposition studied by atomic force microscopy and light scattering [J]. Macromolecules,2001,34(11): 3748-3756.
[27]Kim J H, Kim S C. Effect of PEO grafts on the surface properties of PEO-Grafted PU/PS IPNs:AFM study [J]. Macromolecules,2003,36(8):2867-2872.
[28]Jiang Y, Yan D D, Gao X, et al. Lamellar branching of poly(bisphenol A-co-decane) spherulites at different temperatures studied by high-temperature AFM [J]. Macromolecules, 2003,36(10):3652-3655.
[29]Gibson C W, Thomson N H, Abrams W R, et al. Nested genes:Biological implications and use of AFM for analysis [J]. Gene,2005,350(1):15-23.
[30]Muller D J, Fotiadis D, Scheuring S, et al. Electrostatically balanced subnanometer imaging of biological specimens by atomic force microscope [J]. Biophysical Journal,1999,76(2): 1101-1111.
[31]Jacoboni I, Valdre U, Mori G, et al. Hyaluronic acid by atomic force microscopy [J]. Journal of Structural Biology,1999,126(1):52-58.
[32]Farina M, Schemmel A, Weissmuller G, et al. Atomic force microscopy study of tooth surfaces [J]. Journal of Structural Biology,1999,125(1):39-49.
[33]Martin Y, Williams C C, Wickramasinghe H K. Atomic force microscope-force mapping and profiling on a sub 100-A scale [J]. Journal of Applied Physics,1987,61(10): 4723-4729.
[34]Jalili N, Laxminarayana K. A review of atomic force microscopy imaging systems: application to molecular metrology and biological sciences [J]. Mechatronics,2004,14(8): 907-945.
[35]Janshoff A, Neitzert M, Oberdorfer Y, et al. Force spectroscopy of molecular systems-Single molecule spectroscopy of polymers and biomolecules [J]. Angewandte Chemie-International Edition,2000,39(18):3213-3237.
[36]Hugel T, Seitz M. The study of molecular interactions by AFM force spectroscopy [J]. Macromolecular Rapid Communications,2001,22(13):989-1016.
[37]崔树勋.高分子界面吸附的单分子力谱研究[D].吉林大学,2004.
[38]Merkel R, Nassoy P, Leung A, et al. Energy landscapes of receptor-ligand bonds explored with dynamic force spectroscopy [J]. Nature,1999,397(6714):50-53.
[39]Riener C K, Stroh C M, Ebner A, et al. Simple test system for single molecule recognition force microscopy [J]. Analytica Chimica Acta,2003,479(1):59-75.
[40]Evans E, Ritchie K. Dynamic strength of molecular adhesion bonds [J]. Biophysical Journal,1997,72(4):1541-1555.
[41]Schmitt L, Ludwig M, Gaub H E, et al. A metal-chelating microscopy tip as a new toolbox for single-molecule experiments by atomic force microscopy [J]. Biophysical Journal,2000, 78(6):3275-3285.
[42]Tang J, Krajcikova D, Zhu R, et al. Atomic force microscopy imaging and single molecule recognition force spectroscopy of coat proteins on the surface of Bacillus subtilis spore [J]. Journal of Molecular Recognition,2007,20(6):483-489.
[43]Alessandrini A, Facci P. AFM:a versatile tool in biophysics [J]. Measurement Science & Technology,2005,16(6):R65-R92.
[44]徐永春,师晓丽,方晓红.原子力显微镜单分子力谱研究生物分子间相互作用[J].生命科学,2008,(1)：39-45.
[45]Cappella B, Dietler G. Force-distance curves by atomic force microscopy [J]. Surface Science Reports,1999,34(1-3):1-104.
[46]Liu W, Parpura V. Single molecule probing of SNARE proteins by atomic force microscopy [M]. Mechanisms of Exocytosis.2009:113-120.
[47]Zhang W, Zhang X. Single molecule mechanochemistry of macromolecules [J]. Progress in Polymer Science,2003,28(8):1271-1295.
[48]杨柳.原子力显微镜用于细胞表面结构与性质的研究以及单碱基错配的检测[D].湖南大学,2008.
[49]Li H B, Liu B B, Zhang X, et al. Single-molecule force spectroscopy on poly(acrylic acid) by AFM [J]. Langmuir,1999,15(6):2120-2124.
[50]Hugel T, Grosholz M, Clausen-Schaumann H, et al. Elasticity of single polyelectrolyte chains and their desorption from solid supports studied by AFM based single molecule force spectroscopy [J]. Macromolecules,2001,34(4):1039-1047.
[51]Rief M, Oesterhelt F, Heymann B, et al. Single molecule force spectroscopy on polysaccharides by atomic force microscopy [J]. Science,1997,275(5304):1295-1297.
[52]Norman R S, Frontera-Suau R, Morris P J. Variability in pseudomonas aeruginosa lipopolysaccharide expression during crude oil degradation [J]. Applied and Environment Microbiology,2002,68(10):5096-5103.
[53]Tollersrud T, Berge T, Andersen S R, et al. Imaging the surface of staphylococcus aureus by atomic force microscopy [J]. APMIS,2001,109(7-8):541-545.
[54]Liu C J, Shi W Q, Cui S X, et al. Force spectroscopy of polymers:Beyond single chain mechanics [J]. Current Opinion in Solid State & Materials Science,2005,9(3):140-148.
[55]Bolshakova a V, Kiselyova O I, Filonov a S, et al. Comparative studies of bacteria with an atomic force microscopy operating in different modes [J]. Ultramicroscopy,2001,86(1-2): 121-128.
[56]Hinterdorfer P, Baumgartner W, Gruber H J, et al. Detection and localization of individual antibody-antigen recognition events by atomic force microscopy [J]. Proceedings of the National Academy of Sciences,1996,93(8):3477-3481.
[57]吕正检.基于计算机模拟、自组装和力谱技术的蛋白质分子间相互作用研究[D].重庆大学,2010.
[58]Tang J, Ebner A, Ilk N, et al. High-affinity tags fused to S-Layer proteins probed by atomic force microscopy [J]. Langmuir,2007,24(4):1324-1329.
[59]鲁哲学,张志凌,庞代文.原子力显微镜技术及其在细胞生物学中的应用[J].科学通报,2005,(12)：1161-1166.
[60]张冬仙.原子力显微术的新方法研究及新型原子力显微镜系统研制[D].浙江大学,2004.
[61]Rattner J B, Lin C C. Radial Loops and helical-coils coexist in metaphase chromosomes [J]. Cell,1985,42(1):291-296.
[62]Mcmaster T J, Winfield M O, Baker a A, et al. Chromosome classification by atomic force microscopy volume measurement [J]. Journal of Vacuum Science & Technology B: Microelectronics and Nanometer Structures,1996,14(2):1438-1443.
[63]Murakami M, Minamihisamatsu M, Sato K, et al. Structural analysis of heavy ion radiation-induced chromosome aberrations by atomic force microscopy [J]. Journal of Biochemical and Biophysical Methods,2001,48(3):293-301.
[64]Butt H J, Wolff E K, Gould S a C, et al. Imaging cells with the atomic force microscope [J]. Journal of Structural Biology,1990,105(1-3):54-61.
[65]Lister T E, Pinhero P J. In vivo atomic force microscopy of surface proteins on deinococcus radiodurans [J]. Langmuir,2001,17(9):2624-2628.
[66]Dufrene Y F, Boonaert C J P, Gerin P A, et al. Direct probing of the surface ultrastructure and molecular interactions of dormant and germinating spores of phanerochaete chrysosporium [J]. Journal of Bacteriology,1999,181(17):5350-5354.
[67]Jena B P, Schneider S W, Geibel J P, et al. Gi regulation of secretory vesicle swelling examined by atomic force microscopy [J]. Proceedings of the National Academy of Sciences,1997,94(24):13317-13322.
[68]Amro N A, Kotra L P, Wadu-Mesthrige K, et al. High-resolution atomic force microscopy studies of the Escherichia coli outer membrane:Structural basis for permeability [J]. Langmuir,2000,16(6):2789-2796.
[69]Schneider S W, Pagel P, Rotsch C, et al. Volume dynamics in migrating epithelial cells measured with atomic force microscopy [J]. Pflugers Archiv-European Journal of Physiology,2000,439(3):297-303.
[70]Kawaura C, Noguchi A, Furuno T, et al. Atomic force microscopy for studying gene transfection mediated by cationic liposomes with a cationic cholesterol derivative [J]. Febs Letters,1998,421(1):69-72.
[71]Girasole M, Cricenti A, Generosi R, et al. Atomic force microscopy detects transient frictional contrasts in apoptotic cells induced by deprivation of interleukin-3 [J]. Applied Physics Letters,2001,78(8):1143-1145.
[72]Butt H J, Downing K H, Hansma P K. Imaging the membrane protein bacteriorhodopsin with the atomic force microscope [J]. Biophysical Journal,1990,58(6):1473-1480.
[73]Yang J, Tamm L K, Tillack T W, et al. New approach for atomic force microscopy of membrane proteins:the imaging of cholera toxin [J]. Journal of Molecular Biology,1993, 229(2):286-290.
[74]Schneider S W, Larmer J, Henderson R M, et al. Molecular weights of individual proteins correlate with molecular volumes measured by atomic force microscopy [J]. Pflugers Archiv-European Journal of Physiology,1998,435(3):362-367.
[75]Durbin S D, Carlson W E. Lysozyme crystal growth studied by atomic force microscopy [J]. Journal of Crystal Growth,1992,122(1-4):71-79.
[76]Green N H, Allen S, Davies M C, et al. Force sensing and mapping by atomic force microscopy [J]. Trac-Trends in Analytical Chemistry,2002,21(1):64-73.
[77]Takano H, Kenseth J R, Wong S S, et al. Chemical and biochemical analysis using scanning force microscopy [J]. Chemical Reviews,1999,99(10):2845-2890.
[78]Van Der Aa B C, Michel R M, Asther M, et al. Stretching cell surface macromolecules by atomic force microscopy [J]. Langmuir,2001,17(11):3116-3119.
[79]Schneider S W, Egan M E, Jena B P, et al. Continuous detection of extracellular ATP on living cells by using atomic force microscopy [J]. Proceedings of the National Academy of Sciences of the United States of America,1999,96(21):12180-12185.
[80]Stroh C, Wang H, Bash R, et al. Single-molecule recognition imaging-microscopy [J]. Proceedings of the National Academy of Sciences of the United States of America,2004, 101(34):12503-12507.
[81]Ahimou F, Denis F A, Touhami A, et al. Probing microbial cell surface charges by atomic force microscopy [J]. Langmuir,2002,18(25):9937-9941.
[82]Ong Y L, Razatos A, Georgiou G, et al. Adhesion forces between E-coli bacteria and biomaterial surfaces [J]. Langmuir,1999,15(8):2719-2725.
[83]Grandbois M, Dettmann W, Benoit M, et al. Affinity imaging of red blood cells using an atomic force microscope [J]. Journal of Histochemistry and Cytochemistry,2000,48(5): 719-724.
[84]张成林,李建远.蛋白质相互作用的研究方法[J].中外医学研究,2011,(4)：115-117.
[85]邵壮超.基于多分类器组合的蛋白质—蛋白质相互作用位点预测研究[D].西北工业大学,2006.
[86]Pfleger K D G, Eidne K A. Monitoring the formation of dynamic G-protein-coupled receptor-protein complexes in living cells [J]. Biochemical Journal,2005, (385):625-637.
[87]Florin E, Moy V, Gaub H. Adhesion forces between individual ligand-receptor pairs [J]. Science,1994,264(5157):415-417.
[88]Tanaka T, Sasaki T, Amemiya Y, et al. Discrimination of DNA mismatches by direct force measurement for identification of tuna species [J]. Analytica Chimica Acta,2006,561(1-2): 150-155.
[89]Krasnoslobodtsev a V, Shlyakhtenko L S, Lyubchenko Y L. Probing interactions within the synaptic DNA-Sfil complex by AFM force spectroscopy [J]. Journal of Molecular Biology, 2007,365(5):1407-1416.
[90]Kim J S, Jang S, Kim U, et al. AFM studies of inhibition effect in binding of antimicrobial peptide and immune proteins [J]. Langmuir,2007,23(21):10438-10440.
[91]Pfister G, Stroh C M, Perschinka H, et al. Detection of HSP60 on the membrane surface of stressed human endothelial cells by atomic force and confocal microscopy [J]. Journal of Cell Science,2005,118(8):1587-1594.
[92]Rief M, Gautel M, Oesterhelt F, et al. Reversible unfolding of individual titin immunoglobulin domains by AFM [J]. Science,1997,276(5315):1109-1112.
[93]Janovjak H, Kedrov A, Cisneros D A, et al. Imaging and detecting molecular interactions of single transmembrane proteins [J]. Neurobiology of Aging,2006,27(4):546-561.
[94]Brown a E X, Litvinov R I, Discher D E, et al. Forced unfolding of coiled-coils in fibrinogen by single-molecule AFM [J]. Biophysical Journal,2007,92(5):L39-L41.
[95]Vesenka J, Mosher C, Schaus S, et al. Combining optical and atomic force microscopy for life sciences research [J]. Biotechniques,1995,19(2):240-253.
[96]Nishida S, Funabashi Y, Ikai A. Combination of AFM with an objective-type total internal reflection fluorescence microscope (TIRFM) for nanomanipulation of single cells [J]. Ultramicroscopy,2002,91(1-4):269-274.
[97]Joachimsthaler I, Heiderhoff R, Balk L J. A universal scanning-probe-microscope-based hybrid system [J]. Measurement Science & Technology,2003,14(1):87-96.
[98]郁毅刚,徐如祥,蔡颖谦,et al.原子力显微镜在生命科学研究中的应用[J].第一军医大学学报,2005,(2)：143-147.
[99]Kienberger F, Pastushenko V P, Kada G, et al. Static and dynamical properties of single poly(ethylene glycol) molecules investigated by force spectroscopy [J]. Single Molecules, 2000,1(2):123-128.
[100]Hinterdorfer P, Baumgartner W, Gruber H J, et al. Detection and localization of individual antibody-antigen recognition events by atomic force microscopy [J]. Proceedings of the National Academy of Sciences of the United States of America,1996,93(8):3477-3481.
[101]张志强.黑粉菌[J].生物学通报,1965,(3)：26-28.
[102]杨素贞.玉米丝黑穗病和黑粉病的区别及防治措施[J].现代农村科技,2009,(6)：25.
[103]秦红清.玉米制种田瘤黑粉病的发生与防治[J].中国种业,2008,(12)：76.
[104]王辉,斯琴格日乐,马端辉,et al.玉米黑粉菌孢子粉的无机元素和氨基酸含量测定[J].菌物研究,2006,(1)：20-23.
[105]石竹芹.玉米瘤黑粉病的发生与防治[J].现代农村科技,2009,(6)：31.
[106]Kamper J, Kahmann R, Bolker M, et al. Insights from the genome of the biotrophic fungal plant pathogen Ustilago maydis [J]. Nature,2006,444(7115):97-101.
[107]Banuett F, Herskowitz I. Discrete developmental stages during teliospore formation in the corn smut fungus, Ustilago maydis [J]. Development,1996,122(10):2965-2976.
[108]Klose J, De Sa M M, Kronstad J W. Lipid-induced filamentous growth in Ustilago maydis [J]. Molecular Microbiology,2004,52(3):823-835.
[109]黄青春.羧酰胺类杀菌剂的活性及其机理研究进展[J].世界农药,2004,(4)：23-27.
[110]张一宾.芳酰胺类杀菌剂的沿变——从萎锈灵、灭锈胺、氟酰胺到吡噻菌胺、啶酰菌胺[J].世界农药,2007,(1)：1-7.
[111]杨炜,黄大年,王金霞,邹勤.以潮霉素抗性为选择标记的稻瘟病菌原生质体转化[J].遗传学报,1994,(4)：305-312.
[112]Broomfield P L E, Hargreaves J A. A single amino-acid change in the iron-sulphur protein subunit of succinate dehydrogenase confers resistance to carboxin in Ustilago maydis [J]. Current Genetics,1992,22(2):117-121.
[113]Henriques a O, Moran J, Charles P. Structure, Assembly, and Function of the Spore Surface Layers [J]. Annual Review of Microbiology,2007,61(1):555-588.
[114]Kim H, Hahn M, Grabowski P, et al. The Bacillus subtilis spore coat protein interaction network [J]. Molecular Microbiology,2006,59(2):487-502.
[115]Riesenman P J, Nicholson W L. Role of the spore coat layers in Bacillus subtilis spore resistance to hydrogen peroxide, artificial UV-C, UV-B, and solar UV radiation [J]. Applied and Environmental Microbiology,2000,66(2):620-626.
[116]Driks A. The Bacillus spore coat [J]. Phytopathology,2004,94(11):1249-1251.
[117]Takamatsu H, Watabe K. Assembly and genetics of spore protective structures [J]. Cellular and Molecular Life Sciences,2002,59(3):434-444.
[2]Rost B. Marrying structure and genomics [J]. Structure,1998,6(3):259-263.
[3]Shapiro L, Lima C D. The argonne structural genomics workshop:lamaze class for the birth of a new science [J]. Structure,1998,6(3):265-267.
[4]Venter J C, Adams M D, Myers E W, et al. The sequence of the human genome [J]. Science, 2001,291(5507):1304-1351.
[5]Baltimore D. Our genome unveiled [J]. Nature,2001,409(6822):814-816.
[6]潘宪明.结构生物学——21世纪生命科学的重要前沿[J].中国科学基金,1998,(3)：157-160.
[7]邹承鲁.结构生物学的时代已经开始[J].科技导报,1995,(4)：7-11.
[8]Binnig G, Quate C F, Gerber C. Atomic force microscope [J]. Physical Review Letters, 1986,56(9):930-933.
[9]白春礼.扫描隧道显微术及其应用[M].1992.
[10]Weisenhorn a L, Hansma P K, Albrecht T R, et al. Forces in atomic force microscope in air and water [J]. Applied Physics Letters,1989,54(26):2651-2653.
[11]谭琰.原子力显微镜对生物大分子的研究[D].湘潭大学,2006.
[12]牟旭东.新型扫描力显微镜的研究[D].1998.
[13]陈伟.轻敲模式原子力显微镜的优化研究[D].浙江大学,2002.
[14]Binnig G, Rohrer H, Gerber C, et al. Surface studies by scanning tunneling microscopy [J]. Physical Review Letters,1982,49(1):57.
[15]程舒雯.扫描隧道显微镜性能优化及实用化研究[D].浙江大学,2003.
[16]马尚行.扫描隧道显微镜的新方法研究和系统研制[D].浙江大学,2005.
[17]Allison D P, Hinterdorfer P, Han W. Biomolecular force measurements and the atomic force microscope [J]. Current Opinion in Biotechnology,2002,13(1):47-51.
[18]张德添,何昆,张飒,et al.原子力显微镜发展近况及其应用[J].现代仪器,2002,(3)：6-9.
[19]白春礼.扫描力显微术[M].北京；科学出版社.2000：8-9.
[20]施春陵,蒋建清.原子力显微镜(AFM)在材料性能分析中的应用[J].江苏冶金,2005,(1)：7-9.
[21]鲍俊杰,高明志,周海峰, et al.原子力显微镜在聚氨酯材料性能分析中的应用[J].弹性体,2006,(3)：53-57.
[22]李琦,郑琪,丁焰,et al.原子力显微镜在染色体研究中的应用[J].遗传,2009,(6)：573-580.
[23]程敏熙.STM的同胞兄弟--原子力显微镜(AFM) [J]大学物理,2000,19(9)：5.
[24]Santos N C, Castanho M. An overview of the biophysical applications of atomic force microscopy [J]. Biophysical Chemistry,2004,107(2):133-149.
[25]Maurice P, Forsythe J, Hersman L, et al. Application of atomic-force microscopy to studies of microbial interactions with hydrous Fe(Ⅲ)-oxides [J]. Chemical Geology,1996, 132(1-4):33-43.
[26]Cabral J T, Higgins J S, Mcleish T C B, et al. Bulk spinodal decomposition studied by atomic force microscopy and light scattering [J]. Macromolecules,2001,34(11): 3748-3756.
[27]Kim J H, Kim S C. Effect of PEO grafts on the surface properties of PEO-Grafted PU/PS IPNs:AFM study [J]. Macromolecules,2003,36(8):2867-2872.
[28]Jiang Y, Yan D D, Gao X, et al. Lamellar branching of poly(bisphenol A-co-decane) spherulites at different temperatures studied by high-temperature AFM [J]. Macromolecules, 2003,36(10):3652-3655.
[29]Gibson C W, Thomson N H, Abrams W R, et al. Nested genes:Biological implications and use of AFM for analysis [J]. Gene,2005,350(1):15-23.
[30]Muller D J, Fotiadis D, Scheuring S, et al. Electrostatically balanced subnanometer imaging of biological specimens by atomic force microscope [J]. Biophysical Journal,1999,76(2): 1101-1111.
[31]Jacoboni I, Valdre U, Mori G, et al. Hyaluronic acid by atomic force microscopy [J]. Journal of Structural Biology,1999,126(1):52-58.
[32]Farina M, Schemmel A, Weissmuller G, et al. Atomic force microscopy study of tooth surfaces [J]. Journal of Structural Biology,1999,125(1):39-49.
[33]Martin Y, Williams C C, Wickramasinghe H K. Atomic force microscope-force mapping and profiling on a sub 100-A scale [J]. Journal of Applied Physics,1987,61(10): 4723-4729.
[34]Jalili N, Laxminarayana K. A review of atomic force microscopy imaging systems: application to molecular metrology and biological sciences [J]. Mechatronics,2004,14(8): 907-945.
[35]Janshoff A, Neitzert M, Oberdorfer Y, et al. Force spectroscopy of molecular systems-Single molecule spectroscopy of polymers and biomolecules [J]. Angewandte Chemie-International Edition,2000,39(18):3213-3237.
[36]Hugel T, Seitz M. The study of molecular interactions by AFM force spectroscopy [J]. Macromolecular Rapid Communications,2001,22(13):989-1016.
[37]崔树勋.高分子界面吸附的单分子力谱研究[D].吉林大学,2004.
[38]Merkel R, Nassoy P, Leung A, et al. Energy landscapes of receptor-ligand bonds explored with dynamic force spectroscopy [J]. Nature,1999,397(6714):50-53.
[39]Riener C K, Stroh C M, Ebner A, et al. Simple test system for single molecule recognition force microscopy [J]. Analytica Chimica Acta,2003,479(1):59-75.
[40]Evans E, Ritchie K. Dynamic strength of molecular adhesion bonds [J]. Biophysical Journal,1997,72(4):1541-1555.
[41]Schmitt L, Ludwig M, Gaub H E, et al. A metal-chelating microscopy tip as a new toolbox for single-molecule experiments by atomic force microscopy [J]. Biophysical Journal,2000, 78(6):3275-3285.
[42]Tang J, Krajcikova D, Zhu R, et al. Atomic force microscopy imaging and single molecule recognition force spectroscopy of coat proteins on the surface of Bacillus subtilis spore [J]. Journal of Molecular Recognition,2007,20(6):483-489.
[43]Alessandrini A, Facci P. AFM:a versatile tool in biophysics [J]. Measurement Science & Technology,2005,16(6):R65-R92.
[44]徐永春,师晓丽,方晓红.原子力显微镜单分子力谱研究生物分子间相互作用[J].生命科学,2008,(1)：39-45.
[45]Cappella B, Dietler G. Force-distance curves by atomic force microscopy [J]. Surface Science Reports,1999,34(1-3):1-104.
[46]Liu W, Parpura V. Single molecule probing of SNARE proteins by atomic force microscopy [M]. Mechanisms of Exocytosis.2009:113-120.
[47]Zhang W, Zhang X. Single molecule mechanochemistry of macromolecules [J]. Progress in Polymer Science,2003,28(8):1271-1295.
[48]杨柳.原子力显微镜用于细胞表面结构与性质的研究以及单碱基错配的检测[D].湖南大学,2008.
[49]Li H B, Liu B B, Zhang X, et al. Single-molecule force spectroscopy on poly(acrylic acid) by AFM [J]. Langmuir,1999,15(6):2120-2124.
[50]Hugel T, Grosholz M, Clausen-Schaumann H, et al. Elasticity of single polyelectrolyte chains and their desorption from solid supports studied by AFM based single molecule force spectroscopy [J]. Macromolecules,2001,34(4):1039-1047.
[51]Rief M, Oesterhelt F, Heymann B, et al. Single molecule force spectroscopy on polysaccharides by atomic force microscopy [J]. Science,1997,275(5304):1295-1297.
[52]Norman R S, Frontera-Suau R, Morris P J. Variability in pseudomonas aeruginosa lipopolysaccharide expression during crude oil degradation [J]. Applied and Environment Microbiology,2002,68(10):5096-5103.
[53]Tollersrud T, Berge T, Andersen S R, et al. Imaging the surface of staphylococcus aureus by atomic force microscopy [J]. APMIS,2001,109(7-8):541-545.
[54]Liu C J, Shi W Q, Cui S X, et al. Force spectroscopy of polymers:Beyond single chain mechanics [J]. Current Opinion in Solid State & Materials Science,2005,9(3):140-148.
[55]Bolshakova a V, Kiselyova O I, Filonov a S, et al. Comparative studies of bacteria with an atomic force microscopy operating in different modes [J]. Ultramicroscopy,2001,86(1-2): 121-128.
[56]Hinterdorfer P, Baumgartner W, Gruber H J, et al. Detection and localization of individual antibody-antigen recognition events by atomic force microscopy [J]. Proceedings of the National Academy of Sciences,1996,93(8):3477-3481.
[57]吕正检.基于计算机模拟、自组装和力谱技术的蛋白质分子间相互作用研究[D].重庆大学,2010.
[58]Tang J, Ebner A, Ilk N, et al. High-affinity tags fused to S-Layer proteins probed by atomic force microscopy [J]. Langmuir,2007,24(4):1324-1329.
[59]鲁哲学,张志凌,庞代文.原子力显微镜技术及其在细胞生物学中的应用[J].科学通报,2005,(12)：1161-1166.
[60]张冬仙.原子力显微术的新方法研究及新型原子力显微镜系统研制[D].浙江大学,2004.
[61]Rattner J B, Lin C C. Radial Loops and helical-coils coexist in metaphase chromosomes [J]. Cell,1985,42(1):291-296.
[62]Mcmaster T J, Winfield M O, Baker a A, et al. Chromosome classification by atomic force microscopy volume measurement [J]. Journal of Vacuum Science & Technology B: Microelectronics and Nanometer Structures,1996,14(2):1438-1443.
[63]Murakami M, Minamihisamatsu M, Sato K, et al. Structural analysis of heavy ion radiation-induced chromosome aberrations by atomic force microscopy [J]. Journal of Biochemical and Biophysical Methods,2001,48(3):293-301.
[64]Butt H J, Wolff E K, Gould S a C, et al. Imaging cells with the atomic force microscope [J]. Journal of Structural Biology,1990,105(1-3):54-61.
[65]Lister T E, Pinhero P J. In vivo atomic force microscopy of surface proteins on deinococcus radiodurans [J]. Langmuir,2001,17(9):2624-2628.
[66]Dufrene Y F, Boonaert C J P, Gerin P A, et al. Direct probing of the surface ultrastructure and molecular interactions of dormant and germinating spores of phanerochaete chrysosporium [J]. Journal of Bacteriology,1999,181(17):5350-5354.
[67]Jena B P, Schneider S W, Geibel J P, et al. Gi regulation of secretory vesicle swelling examined by atomic force microscopy [J]. Proceedings of the National Academy of Sciences,1997,94(24):13317-13322.
[68]Amro N A, Kotra L P, Wadu-Mesthrige K, et al. High-resolution atomic force microscopy studies of the Escherichia coli outer membrane:Structural basis for permeability [J]. Langmuir,2000,16(6):2789-2796.
[69]Schneider S W, Pagel P, Rotsch C, et al. Volume dynamics in migrating epithelial cells measured with atomic force microscopy [J]. Pflugers Archiv-European Journal of Physiology,2000,439(3):297-303.
[70]Kawaura C, Noguchi A, Furuno T, et al. Atomic force microscopy for studying gene transfection mediated by cationic liposomes with a cationic cholesterol derivative [J]. Febs Letters,1998,421(1):69-72.
[71]Girasole M, Cricenti A, Generosi R, et al. Atomic force microscopy detects transient frictional contrasts in apoptotic cells induced by deprivation of interleukin-3 [J]. Applied Physics Letters,2001,78(8):1143-1145.
[72]Butt H J, Downing K H, Hansma P K. Imaging the membrane protein bacteriorhodopsin with the atomic force microscope [J]. Biophysical Journal,1990,58(6):1473-1480.
[73]Yang J, Tamm L K, Tillack T W, et al. New approach for atomic force microscopy of membrane proteins:the imaging of cholera toxin [J]. Journal of Molecular Biology,1993, 229(2):286-290.
[74]Schneider S W, Larmer J, Henderson R M, et al. Molecular weights of individual proteins correlate with molecular volumes measured by atomic force microscopy [J]. Pflugers Archiv-European Journal of Physiology,1998,435(3):362-367.
[75]Durbin S D, Carlson W E. Lysozyme crystal growth studied by atomic force microscopy [J]. Journal of Crystal Growth,1992,122(1-4):71-79.
[76]Green N H, Allen S, Davies M C, et al. Force sensing and mapping by atomic force microscopy [J]. Trac-Trends in Analytical Chemistry,2002,21(1):64-73.
[77]Takano H, Kenseth J R, Wong S S, et al. Chemical and biochemical analysis using scanning force microscopy [J]. Chemical Reviews,1999,99(10):2845-2890.
[78]Van Der Aa B C, Michel R M, Asther M, et al. Stretching cell surface macromolecules by atomic force microscopy [J]. Langmuir,2001,17(11):3116-3119.
[79]Schneider S W, Egan M E, Jena B P, et al. Continuous detection of extracellular ATP on living cells by using atomic force microscopy [J]. Proceedings of the National Academy of Sciences of the United States of America,1999,96(21):12180-12185.
[80]Stroh C, Wang H, Bash R, et al. Single-molecule recognition imaging-microscopy [J]. Proceedings of the National Academy of Sciences of the United States of America,2004, 101(34):12503-12507.
[81]Ahimou F, Denis F A, Touhami A, et al. Probing microbial cell surface charges by atomic force microscopy [J]. Langmuir,2002,18(25):9937-9941.
[82]Ong Y L, Razatos A, Georgiou G, et al. Adhesion forces between E-coli bacteria and biomaterial surfaces [J]. Langmuir,1999,15(8):2719-2725.
[83]Grandbois M, Dettmann W, Benoit M, et al. Affinity imaging of red blood cells using an atomic force microscope [J]. Journal of Histochemistry and Cytochemistry,2000,48(5): 719-724.
[84]张成林,李建远.蛋白质相互作用的研究方法[J].中外医学研究,2011,(4)：115-117.
[85]邵壮超.基于多分类器组合的蛋白质—蛋白质相互作用位点预测研究[D].西北工业大学,2006.
[86]Pfleger K D G, Eidne K A. Monitoring the formation of dynamic G-protein-coupled receptor-protein complexes in living cells [J]. Biochemical Journal,2005, (385):625-637.
[87]Florin E, Moy V, Gaub H. Adhesion forces between individual ligand-receptor pairs [J]. Science,1994,264(5157):415-417.
[88]Tanaka T, Sasaki T, Amemiya Y, et al. Discrimination of DNA mismatches by direct force measurement for identification of tuna species [J]. Analytica Chimica Acta,2006,561(1-2): 150-155.
[89]Krasnoslobodtsev a V, Shlyakhtenko L S, Lyubchenko Y L. Probing interactions within the synaptic DNA-Sfil complex by AFM force spectroscopy [J]. Journal of Molecular Biology, 2007,365(5):1407-1416.
[90]Kim J S, Jang S, Kim U, et al. AFM studies of inhibition effect in binding of antimicrobial peptide and immune proteins [J]. Langmuir,2007,23(21):10438-10440.
[91]Pfister G, Stroh C M, Perschinka H, et al. Detection of HSP60 on the membrane surface of stressed human endothelial cells by atomic force and confocal microscopy [J]. Journal of Cell Science,2005,118(8):1587-1594.
[92]Rief M, Gautel M, Oesterhelt F, et al. Reversible unfolding of individual titin immunoglobulin domains by AFM [J]. Science,1997,276(5315):1109-1112.
[93]Janovjak H, Kedrov A, Cisneros D A, et al. Imaging and detecting molecular interactions of single transmembrane proteins [J]. Neurobiology of Aging,2006,27(4):546-561.
[94]Brown a E X, Litvinov R I, Discher D E, et al. Forced unfolding of coiled-coils in fibrinogen by single-molecule AFM [J]. Biophysical Journal,2007,92(5):L39-L41.
[95]Vesenka J, Mosher C, Schaus S, et al. Combining optical and atomic force microscopy for life sciences research [J]. Biotechniques,1995,19(2):240-253.
[96]Nishida S, Funabashi Y, Ikai A. Combination of AFM with an objective-type total internal reflection fluorescence microscope (TIRFM) for nanomanipulation of single cells [J]. Ultramicroscopy,2002,91(1-4):269-274.
[97]Joachimsthaler I, Heiderhoff R, Balk L J. A universal scanning-probe-microscope-based hybrid system [J]. Measurement Science & Technology,2003,14(1):87-96.
[98]郁毅刚,徐如祥,蔡颖谦,et al.原子力显微镜在生命科学研究中的应用[J].第一军医大学学报,2005,(2)：143-147.
[99]Kienberger F, Pastushenko V P, Kada G, et al. Static and dynamical properties of single poly(ethylene glycol) molecules investigated by force spectroscopy [J]. Single Molecules, 2000,1(2):123-128.
[100]Hinterdorfer P, Baumgartner W, Gruber H J, et al. Detection and localization of individual antibody-antigen recognition events by atomic force microscopy [J]. Proceedings of the National Academy of Sciences of the United States of America,1996,93(8):3477-3481.
[101]张志强.黑粉菌[J].生物学通报,1965,(3)：26-28.
[102]杨素贞.玉米丝黑穗病和黑粉病的区别及防治措施[J].现代农村科技,2009,(6)：25.
[103]秦红清.玉米制种田瘤黑粉病的发生与防治[J].中国种业,2008,(12)：76.
[104]王辉,斯琴格日乐,马端辉,et al.玉米黑粉菌孢子粉的无机元素和氨基酸含量测定[J].菌物研究,2006,(1)：20-23.
[105]石竹芹.玉米瘤黑粉病的发生与防治[J].现代农村科技,2009,(6)：31.
[106]Kamper J, Kahmann R, Bolker M, et al. Insights from the genome of the biotrophic fungal plant pathogen Ustilago maydis [J]. Nature,2006,444(7115):97-101.
[107]Banuett F, Herskowitz I. Discrete developmental stages during teliospore formation in the corn smut fungus, Ustilago maydis [J]. Development,1996,122(10):2965-2976.
[108]Klose J, De Sa M M, Kronstad J W. Lipid-induced filamentous growth in Ustilago maydis [J]. Molecular Microbiology,2004,52(3):823-835.
[109]黄青春.羧酰胺类杀菌剂的活性及其机理研究进展[J].世界农药,2004,(4)：23-27.
[110]张一宾.芳酰胺类杀菌剂的沿变——从萎锈灵、灭锈胺、氟酰胺到吡噻菌胺、啶酰菌胺[J].世界农药,2007,(1)：1-7.
[111]杨炜,黄大年,王金霞,邹勤.以潮霉素抗性为选择标记的稻瘟病菌原生质体转化[J].遗传学报,1994,(4)：305-312.
[112]Broomfield P L E, Hargreaves J A. A single amino-acid change in the iron-sulphur protein subunit of succinate dehydrogenase confers resistance to carboxin in Ustilago maydis [J]. Current Genetics,1992,22(2):117-121.
[113]Henriques a O, Moran J, Charles P. Structure, Assembly, and Function of the Spore Surface Layers [J]. Annual Review of Microbiology,2007,61(1):555-588.
[114]Kim H, Hahn M, Grabowski P, et al. The Bacillus subtilis spore coat protein interaction network [J]. Molecular Microbiology,2006,59(2):487-502.
[115]Riesenman P J, Nicholson W L. Role of the spore coat layers in Bacillus subtilis spore resistance to hydrogen peroxide, artificial UV-C, UV-B, and solar UV radiation [J]. Applied and Environmental Microbiology,2000,66(2):620-626.
[116]Driks A. The Bacillus spore coat [J]. Phytopathology,2004,94(11):1249-1251.
[117]Takamatsu H, Watabe K. Assembly and genetics of spore protective structures [J]. Cellular and Molecular Life Sciences,2002,59(3):434-444.