• 文献检索
  • 文档翻译
  • 深度研究
  • 学术资讯
  • Suppr Zotero 插件Zotero 插件
  • 邀请有礼
  • 套餐&价格
  • 历史记录
应用&插件
Suppr Zotero 插件Zotero 插件浏览器插件Mac 客户端Windows 客户端微信小程序
定价
高级版会员购买积分包购买API积分包
服务
文献检索文档翻译深度研究API 文档MCP 服务
关于我们
关于 Suppr公司介绍联系我们用户协议隐私条款
关注我们

Suppr 超能文献

核心技术专利：CN118964589B侵权必究
粤ICP备2023148730 号-1Suppr @ 2026

文献检索

告别复杂PubMed语法，用中文像聊天一样搜索，搜遍4000万医学文献。AI智能推荐，让科研检索更轻松。

立即免费搜索

文件翻译

保留排版，准确专业，支持PDF/Word/PPT等文件格式，支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述，25分钟生成高质量综述，智能提取关键信息，辅助科研写作。

立即免费体验

通过化学修饰检测DNA中的单碱基对错配，随后在15%聚丙烯酰胺凝胶中进行电泳。

Detection of single base-pair mismatches in DNA by chemical modification followed by electrophoresis in 15% polyacrylamide gel.

作者信息

Novack D F, Casna N J, Fischer S G, Ford J P

出版信息

Proc Natl Acad Sci U S A. 1986 Feb;83(3):586-90. doi: 10.1073/pnas.83.3.586.

DOI:10.1073/pnas.83.3.586
PMID:3003741
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC322908/
Abstract

We have developed a method for distinguishing fragments of DNA that contain single-base mismatches from their perfectly paired homologues. Single-stranded regions within a duplex fragment are accessible to 1-cyclohexyl-3-(2-[4-(4-methyl)morpholinyl]ethyl)carbodiimide, which reacts with unpaired guanidylate and thymidylate residues in DNA. Intact linear duplex DNA molecules do not react with carbodiimide, whereas DNA molecules containing single-base mismatches react quantitatively. After carbodiimide reaction, the DNA molecules are electrophoresed in high-percentage polyacrylamide gels so that modified and unmodified fragments can be resolved. Application of this technique should make it possible to locate and purify DNA fragments that exhibit sequence differences from those that do not; these might be used to signal phenotypic variation as well as to diagnose inherited disease.

摘要

我们已经开发出一种方法，用于区分含有单碱基错配的DNA片段与其完全配对的同源片段。双链片段中的单链区域可被1-环己基-3-(2-[4-(4-甲基)吗啉基]乙基)碳二亚胺作用，该试剂会与DNA中未配对的鸟苷酸和胸苷酸残基发生反应。完整的线性双链DNA分子不会与碳二亚胺反应，而含有单碱基错配的DNA分子则会发生定量反应。碳二亚胺反应后，将DNA分子在高百分比聚丙烯酰胺凝胶中进行电泳，以便分离修饰和未修饰的片段。应用这项技术应该能够定位和纯化那些与无序列差异的片段表现出序列不同的DNA片段；这些片段可用于指示表型变异以及诊断遗传性疾病。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/1c8c21c50103/pnas00307-0070-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/4028b1e5a177/pnas00307-0069-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/5b7f7acb8726/pnas00307-0069-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/8cde1a85f9b7/pnas00307-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/d4c836d1a7f4/pnas00307-0070-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/1c8c21c50103/pnas00307-0070-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/4028b1e5a177/pnas00307-0069-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/5b7f7acb8726/pnas00307-0069-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/8cde1a85f9b7/pnas00307-0070-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/d4c836d1a7f4/pnas00307-0070-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/b85f/322908/1c8c21c50103/pnas00307-0070-c.jpg

相似文献

1
Detection of single base-pair mismatches in DNA by chemical modification followed by electrophoresis in 15% polyacrylamide gel.通过化学修饰检测DNA中的单碱基对错配，随后在15%聚丙烯酰胺凝胶中进行电泳。
Proc Natl Acad Sci U S A. 1986 Feb;83(3):586-90. doi: 10.1073/pnas.83.3.586.
2
Immunoassays for carbodiimide modified DNA-detection of unpairing transitions in supercoiled ColE1 DNA.用于碳二亚胺修饰DNA的免疫测定——超螺旋ColE1 DNA中解链转变的检测
Nucleic Acids Res. 1989 Dec 11;17(23):9957-77. doi: 10.1093/nar/17.23.9957.
3
Detection of single-base mutations by reaction of DNA heteroduplexes with a water-soluble carbodiimide followed by primer extension: application to products from the polymerase chain reaction.通过DNA异源双链体与水溶性碳二亚胺反应后进行引物延伸来检测单碱基突变：应用于聚合酶链反应产物
Nucleic Acids Res. 1990 Jul 11;18(13):3933-9. doi: 10.1093/nar/18.13.3933.
4
Binding of DNA to albumin and transferrin modified by treatment with water-soluble carbodiimides.DNA与经水溶性碳二亚胺处理修饰的白蛋白和转铁蛋白的结合。
Biochem Pharmacol. 1986 Apr 15;35(8):1249-57. doi: 10.1016/0006-2952(86)90267-4.
5
Reactivity of ferrocenylcarbodiimide with DNA duplex containing single-mismatched base pairs.二茂铁基碳二亚胺与含有单错配碱基对的DNA双链体的反应活性。
Anal Sci. 2007 Jun;23(6):645-9. doi: 10.2116/analsci.23.645.
6
Modification of a ribonuclease from Rhizopus sp. with 1-cyclohexyl-3-(2-morpholinyl-(4)-ethyl)carbodiimide p-toluenesulfonate.用对甲苯磺酸1-环己基-3-(2-吗啉基-(4)-乙基)碳二亚胺修饰来自根霉属的一种核糖核酸酶。
J Biochem. 1985 Jul;98(1):125-32. doi: 10.1093/oxfordjournals.jbchem.a135250.
7
Detection and location of single-base mutations in large DNA fragments by immunomicroscopy.通过免疫显微镜检测和定位大DNA片段中的单碱基突变
Genomics. 1989 May;4(4):530-8. doi: 10.1016/0888-7543(89)90276-0.
8
A highly sensitive technique for staining DNA and RNA in polyacrylamide gels using silver.一种使用银对聚丙烯酰胺凝胶中的DNA和RNA进行染色的高灵敏度技术。
J Biochem Biophys Methods. 1981 Oct;5(4):219-28. doi: 10.1016/0165-022x(81)90046-4.
9
A new packing for separation of DNA restriction fragments by high performance liquid chromatography.一种用于通过高效液相色谱法分离DNA限制性片段的新型填料。
J Biochem. 1984 Jan;95(1):83-6. doi: 10.1093/oxfordjournals.jbchem.a134606.
10
Separation of large DNA restriction fragments by high-performance gel filtration on TSKgel DNA-PW.使用TSKgel DNA-PW通过高效凝胶过滤分离大的DNA限制性片段。
J Chromatogr. 1985 Mar 1;320(2):440-4. doi: 10.1016/s0021-9673(01)90526-6.

引用本文的文献

1
Conformational changes of the phenyl and naphthyl isocyanate-DNA adducts during DNA replication and by minor groove binding molecules.苯异氰酸酯-DNA 加合物和萘异氰酸酯-DNA 加合物在 DNA 复制过程中的构象变化及对碱基小沟的结合分子。
Nucleic Acids Res. 2013 Oct;41(18):8581-90. doi: 10.1093/nar/gkt608. Epub 2013 Jul 19.
2
Improving CMC-derivatization of pseudouridine in RNA for mass spectrometric detection.改进用于质谱检测的RNA中假尿苷的化学酶法衍生化
Anal Chim Acta. 2008 Apr 7;612(2):173-81. doi: 10.1016/j.aca.2008.02.026. Epub 2008 Feb 17.
3
A novel procedure for simple and efficient genotyping of single nucleotide polymorphisms by using the Zn2+-cyclen complex.

本文引用的文献

1
Chemical modification studies and the secondary structure of HeLa cell 5.8S rRNA.海拉细胞5.8S核糖体RNA的化学修饰研究及其二级结构
Nucleic Acids Res. 1980 Oct 10;8(19):4521-34. doi: 10.1093/nar/8.19.4521.
2
Oligonucleotide directed mutagenesis of the human beta-globin gene: a general method for producing specific point mutations in cloned DNA.人β-珠蛋白基因的寡核苷酸定向诱变：在克隆DNA中产生特定位点突变的通用方法。
Nucleic Acids Res. 1981 Aug 11;9(15):3647-56. doi: 10.1093/nar/9.15.3647.
3
Possible molecular detent in the DNA structure at regulatory sequences.
一种使用Zn2+-轮环化合物进行单核苷酸多态性简单高效基因分型的新方法。
Nucleic Acids Res. 2002 Nov 15;30(22):e126. doi: 10.1093/nar/gnf126.
4
Conformation sensitive gel electrophoresis for simple and accurate detection of mutations: comparison with denaturing gradient gel electrophoresis and nucleotide sequencing.用于简单准确检测突变的构象敏感凝胶电泳：与变性梯度凝胶电泳和核苷酸测序的比较
Proc Natl Acad Sci U S A. 1998 Feb 17;95(4):1681-5. doi: 10.1073/pnas.95.4.1681.
5
Mutation detection with MutH, MutL, and MutS mismatch repair proteins.利用MutH、MutL和MutS错配修复蛋白进行突变检测。
Proc Natl Acad Sci U S A. 1996 Apr 30;93(9):4374-9. doi: 10.1073/pnas.93.9.4374.
6
Single base pair mutation analysis by PNA directed PCR clamping.通过肽核酸定向聚合酶链式反应钳夹进行单碱基对突变分析。
Nucleic Acids Res. 1993 Nov 25;21(23):5332-6. doi: 10.1093/nar/21.23.5332.
7
Conformation-sensitive gel electrophoresis for rapid detection of single-base differences in double-stranded PCR products and DNA fragments: evidence for solvent-induced bends in DNA heteroduplexes.用于快速检测双链PCR产物和DNA片段中单碱基差异的构象敏感性凝胶电泳：DNA异源双链体中溶剂诱导弯曲的证据
Proc Natl Acad Sci U S A. 1993 Nov 1;90(21):10325-9. doi: 10.1073/pnas.90.21.10325.
8
Genomic analysis I: inheritance units and genetic selection in the rapid discovery of locus linked DNA markers.基因组分析I：在快速发现与基因座连锁的DNA标记中的遗传单位和遗传选择
Nucleic Acids Res. 1986 Sep 25;14(18):7265-83. doi: 10.1093/nar/14.18.7265.
9
Ionizing radiation-induced mutagenesis.电离辐射诱导的诱变作用。
Br J Cancer. 1988 Jan;57(1):6-18. doi: 10.1038/bjc.1988.2.
10
Reactivity of cytosine and thymine in single-base-pair mismatches with hydroxylamine and osmium tetroxide and its application to the study of mutations.单碱基对错配中胞嘧啶和胸腺嘧啶与羟胺和四氧化锇的反应性及其在突变研究中的应用
Proc Natl Acad Sci U S A. 1988 Jun;85(12):4397-401. doi: 10.1073/pnas.85.12.4397.
调控序列处DNA结构中可能存在的分子制动。
J Biomol Struct Dyn. 1983 Oct;1(2):509-21. doi: 10.1080/07391102.1983.10507458.
4
Analysis of the transforming potential of the human H-ras gene by random mutagenesis.通过随机诱变分析人类H-ras基因的转化潜力。
Proc Natl Acad Sci U S A. 1984 Jul;81(13):4008-12. doi: 10.1073/pnas.81.13.4008.
5
alpha 1-antitrypsin deficiency detection by direct analysis of the mutation in the gene.通过对基因中的突变进行直接分析来检测α1-抗胰蛋白酶缺乏症。
Nature. 1983;304(5923):230-4. doi: 10.1038/304230a0.
6
Complete nucleotide sequences of the T24 human bladder carcinoma oncogene and its normal homologue.T24人膀胱癌癌基因及其正常同源基因的完整核苷酸序列。
Nature. 1983 Mar 3;302(5903):33-7. doi: 10.1038/302033a0.
7
A rapid boiling method for the preparation of bacterial plasmids.一种制备细菌质粒的快速煮沸法。
Anal Biochem. 1981 Jun;114(1):193-7. doi: 10.1016/0003-2697(81)90473-5.
8
A versatile primer for DNA sequencing in the M13mp2 cloning system.M13mp2克隆系统中用于DNA测序的通用引物。
Gene. 1980 Jun;10(1):69-73. doi: 10.1016/0378-1119(80)90145-6.
9
Sequence-determined DNA separations.序列决定的DNA分离
Annu Rev Biophys Bioeng. 1984;13:399-423. doi: 10.1146/annurev.bb.13.060184.002151.
10
DNA fragments differing by single base-pair substitutions are separated in denaturing gradient gels: correspondence with melting theory.通过单碱基对替换而不同的DNA片段在变性梯度凝胶中被分离：与解链理论的对应关系。
Proc Natl Acad Sci U S A. 1983 Mar;80(6):1579-83. doi: 10.1073/pnas.80.6.1579.