SCI诞生50周年《自然》揭晓百篇高引论文榜单 - 社区报道 - 最新消息

SCI诞生50周年《自然》揭晓百篇高引论文榜单

来源：侨报网｜ 2014/11/1 8:57:33 ｜浏览：3127 ｜评论：0

为了纪念科学引文索引（SCI）诞生五十周年，Nature委托汤森路透列出了数据库中的百大高引论文。荣登榜首的那篇文章你一定猜不到。

这份百强名单的入围资格是引用次数超过12119。令人意外的是，许多特别知名的论文并未入榜，比如DNA双螺旋结构的确定。而1951年发表的一种蛋白定量方法，狂揽三十多万次引用，成为了最大的赢家。该文章的作者，美国生化学家Oliver Lowry也对此感到困惑。

Rank	Title	Times cited
1	Protein measurement with the folin phenol reagent.	305148
2	Cleavage of structural proteins during the assembly of the head of bacteriophage T4.	213005
3	A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding.	155530
4	DNA sequencing with chain-terminating inhibitors.	65335
5	Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction.	60397
6	Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets：procedure and some applications.	53349
7	Development of the Colle-Salvetti correlation-energy formula into a functional of the electron density.	46702
8	Density-functional thermochemistry. III. The role of exact exchange.	46145
9	A simple method for the isolation and purification of total lipides from animal tissues.	45131
10	Clustal W：improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.	40289
11	Nonparametric estimation from incomplete observations.	38600
12	Basic local alignment search tool.	38380
13	A short history of SHELX.	37978
14	Gapped BLAST and PSI-BLAST：A new generation of protein database search programs.	36410
15	A revised medium for rapid growth and bio assays with tobacco tissue cultures.	36132
16	Generalized gradient approximation made simple.	35405
17	"Mini-mental state"：A practical method for grading cognitive state of patients for clinician.	34532
18	A rapid method of total lipid extraction and purification.	32131
19	Detection of specific sequences among DNA fragments separated by gel-electrophoresis.	31904
20	The neighbor-joining method：A new method for reconstructing phylogenetic trees.	30176
21	Analysis of relative gene expression data using real-time quantitative PCR and the 2（T）（-Delta Delta C）method.	28870
22	Revised effective ionic radii and systematic studies of interatomic distances in halides and chalcogenides.	28658
23	Processing of X-ray diffraction data collected in oscillation mode.	28647
24	Regression models and life-tables.	28439
25	Density-functional exchange-energy approximation with correct asymptotic-behavior.	26475
26	Colorimetric method for determination of sugars and related substances.	25735
27	Use of lead citrate at high pH as an electron-opaque stain in electron microscopy.	24449
28	The CLUSTAL_X Windows interface：Flexible strategies for multiple sequence alignment aided by quality analysis tools.	24098
29	Statistical methods for assessing agreement between two methods of clinical measurement.	23826
30	Reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis.	23642
31	Isolation of biologically-active ribonucleic-acid from sources enriched in ribonuclease.	23435
32	The attractions of proteins for small molecules and ions.	23421
33	The moderator–mediator variable distinction in social psychological-research — conceptual, strategic, and statistical considerations.	23356
34	Self-consistent equations including exchange and correlation effects.	23059
35	Rapid colorimetric assay for cellular growth and survival — application to proliferation and cyto-toxicity assays.	23011
36	Helical microtubules of graphitic carbon.	22899
37	The colorimetric determination of phosphorus.	22690
38	Disc electrophoresis — II. Method and application to human serum proteins.	22074
39	Inhomogeneous electron gas.	21931
40	A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.	21446
41	Confidence limits on phylogenies：an approach using the bootstrap	21373
42	A new generation of Ca2+ indicators with greatly improved fluorescence properties	19561
43	Efficient iterative schemes for ab initio total-energy calculations using a plane-wave basis set.	18856
44	High-resolution 2-dimensional electrophoresis of proteins.	18489
45	MEGA4：Molecular Evolutionary Genetics Analysis（MEGA）software version 4.0.	18286
46	Fuzzy sets.	18203
47	Phase annealing in SHELX-90：direct methods for larger structures.	17728
48	Clinical diagnosis of Alzheimer’s disease：Report of the NINCDS-ADRDA Work Group under the auspices of Department of Health and Human Services Task Force on Alzheimer’s Disease.	17220
49	Special points for Brillouin-zone integrations.	17087
50	Study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid.	17067
51	The CES-D scale：a self-report depression scale for research in the general population.	17055
52	Improved patch-clamp techniques for high-resolution current recording from cells and cell-free membrane patches.	17025
53	A rating scale for depression.	16734
54	An inventory for measuring depression.	16264
55	A simple method for displaying the hydropathic character of a protein.	16059
56	Determination of serum proteins by means of the biuret reaction.	16009
57	Maximum likelihood from incomplete data via EM algorithm.	15993
58	Equation of state calculations by fast computing machines.	15902
59	Controlling the false discovery rate：a practical and powerful approach to multiple testing.	15898
60	Measurement of protein using bicinchoninic acid.	15802
61	The assessment and analysis of handedness：the Edinburgh inventory.	15517
62	Estimation of concentration of low-density lipoprotein cholesterol in plasma, without use of preparative ultracentrifuge.	15469
63	Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.	15160
64	Multiple range and multiple F tests.	15047
65	Electric field effect in atomically thin carbon films.	15022
66	Tissue sulfhydryl groups.	15019
67	Isolation of mononuclear cells and granulocytes from human blood.	14934
68	The measurement of observer agreement for categorical data.	14903
69	Crystallography & NMR system：a new software suite for macromolecular structure determination.	14898
70	Gaussian-basis sets for use in correlated molecular calculations. 1. The atoms boron through neon and hydrogen.	14617
71	PROCHECK：a program to check the stereochemical quality of protein structures.	14462
72	The MOS 36-item short-form health survey（SF-36）：I. Conceptual framework and item selection.	14332
73	A new look at statistical-model identification.	14275
74	Improved M13 phage cloning vectors and host strains — nucleotide-sequences of the M13mp18 and pUC19 vectors	14232
75	A comprehensive set of sequence-analysis programs for the vax.	14226
76	MODELTEST：Testing the model of DNA.	14099
77	From ultrasoft pseudopotentials to the projector augmented-wave method.	14049
78	Use of avidin-biotin-peroxidase complex（ABC）in immunoperoxidase techniques：a comparison between ABC and unlabeled antibody（PAP）procedures.	13881
79	Comparison of simple potential functions for simulating liquid water.	13774
80	The development and use of quantum-mechanical molecular-models. 76. AM1：a new general-purpose quantum-mechanical molecular-model	13718
81	Phosphorus assay in column chromatography.	13523
82	MOLSCRIPT：a program to produce both detailed and schematic plots of protein structures.	13496
83	Van der Waals volumes and radii.	13417
84	A new and rapid colorimetric determination of acetylcholinesterase activity.	13332
85	Projector augmented-wave method.	13330
86	Optimization by simulated annealing.	13293
87	Nitric oxide：physiology, pathophysiology, and pharmacology.	13267
88	An algorithm for least-squares estimation of nonlinear parameters.	13258
89	Efficiency of ab initio total energy calculations for metals and semiconductors using a plane-wave basis set.	13084
90	A low-cost, high-efficiency solar-cell based on dye-sensitized colloidal TiO2 films.	12873
91	A low-viscosity epoxy resin embedding medium for electron microscopy.	12807
92	The Protein Data Bank.	12754
93	Accurate and simple analytic representation of the electron-gas correlation-energy	12748
94	Rapid alkaline extraction procedure for screening recombinant plasmid DNA.	12721
95	Improved methods for building protein models in electron-density maps and the location of errors in these models.	12649
96	Accurate spin-dependent electron liquid correlation energies for local spin-density calculations — a critical analysis.	12583
97	Continuous cultures of fused cells secreting antibody of predefined specificity.	12391
98	Homeostasis model assessment：insulin resistance and beta-cell function from fasting plasma glucose and insulin concentrations in man.	12257
99	Adsorption of gases in multimolecular layers.	12252
100	MrBayes 3：Bayesian phylogenetic inference under mixed models	12209
101	Atherosclerosis — an inflammatory disease.	12119

那么百强榜上还有哪些与生物学有关的关键方法呢？

生物学技术

引文百强主要被蛋白质生化论文所占据。1951年发表的Lowry蛋白定量法遥遥领先，而Bradford蛋白定量法位列第三。排在它们之间的是用于蛋白分析的Laemmli缓冲液。这些技术在细胞和分子生物学文章中被大量引用，它们仍是这些领域最基本的工具之一。

上榜的生物学技术中有两个获得了诺贝尔奖。Frederick Sanger的DNA测序方法位列第四，该方法也为他赢得了1980年的诺贝尔化学奖。Kary Mullis因PCR法获得了1993年的诺贝尔化学奖，这一方法在引文百强中位列63。测序和PCR都是遗传学研究的革命性技术，如今仍然有着旺盛的生命力。

除此之外，上世纪八十年代的一种RNA纯化技术位列第五。该技术实现了低成本的快速RNA提取，受到了广泛的欢迎。在非编码短RNA的研究热潮中，这一技术仍然起着重要的作用。

生物信息学

基因测序的迅猛发展，大大提高了序列分析论文的排名，BLAST就是一个很好的例子。对于研究基因和蛋白功能的研究者来说，BLAST近二十年来绝对算得上是家喻户晓。我们只需要打开一个网页，输入DNA、RNA或蛋白序列，就能在几秒钟内获得各种生物中的相关序列。BLAST的两个版本居然都上了百强榜，分别在12位和14位。

能够一次比对多个序列的Clustal也不甘示弱，ClustalW和ClustalX的排名分别是第10和第28。在Clustal软件的帮助下，人们能研究源自不同生物的序列有怎样的进化关系，看起来不想关的序列存在怎样的匹配，预测DNA或蛋白特定位点发生改变对其功能有何影响。

系统发生

系统发生学主要是研究物种之间的进化关系，这类方法也随着基因组测序的发展而水涨船高。

百强榜的第二十位是neighbor-joining法。这种方法由根井正利（Masatoshi Nei）等人提出，可以根据大量物种之间的进化距离，快速有效的构建进化树。

排在第41位的文章，将统计学工具用到了系统发生研究中。这种称为bootstrap的工具，能够推断进化树不同部分的精确性。

百强榜上的其他生物学相关方法，还包括一些用在生物医学领域的统计学方法，以及晶体结构分析用到的工具。

“如果你想要的是被引用，”耶鲁大学的化学家Peter Moore评论道。“那么开发一种人们需要的方法比探寻宇宙的秘密更容易达到目的。”

原文检索：

Richard Van Noorden,Brendan Maher & Regina Nuzzo. The top 100 papers. Nature, 30 October 2014; doi:10.1038/514550a

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