Nature百大高引论文 榜首竟然是...
2014/11/3 医学生
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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。
据介绍，这份百强名单的入围资格是引用次数超过12,119。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。”
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这里不仅有生命科学领域最前沿的研究、最有趣的知识，还有医药产业最权威的资讯、最火爆的话题，更有一大波神秘活动等着你!生物谷，生物Good!
赛先生微信号：Iscientists(←长按复制搜索)
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