一种用于双链质粒DNA定点诱变的简单高效方法。
A simple and efficient method for site-directed mutagenesis with double-stranded plasmid DNA.
作者信息
Lai D, Zhu X, Pestka S
机构信息
Department of Molecular Genetics and Microbiology, UMDNJ-Robert Wood Johnson Medical School, Piscataway 08854-5635.
出版信息
Nucleic Acids Res. 1993 Aug 25;21(17):3977-80. doi: 10.1093/nar/21.17.3977.
Abstract
A general, simple and efficient method for preparing site-specific mutations in double-stranded plasmid DNA without the need for special plasmids, bacterial strains or reagents is described. Only one synthetic oligonucleotide for each mutation is required, subcloning is unnecessary and a high efficiency of mutation (58-97%) was obtained. If two synthetic oligonucleotide primers are used, two separate mutations can be simultaneously created in a single reaction tube.
摘要
本文描述了一种通用、简单且高效的方法，用于在双链质粒DNA中制备位点特异性突变，无需特殊质粒、细菌菌株或试剂。每个突变仅需一个合成寡核苷酸，无需亚克隆，且获得了较高的突变效率（58-97%）。如果使用两个合成寡核苷酸引物，则可在单个反应管中同时产生两个独立的突变。
相似文献
1
A simple and efficient method for site-directed mutagenesis with double-stranded plasmid DNA.一种用于双链质粒DNA定点诱变的简单高效方法。
Nucleic Acids Res. 1993 Aug 25;21(17):3977-80. doi: 10.1093/nar/21.17.3977.
2
A rapid and versatile site-directed method of mutagenesis for double-stranded plasmid DNA.
Gene. 1988 Sep 30;69(2):325-30. doi: 10.1016/0378-1119(88)90442-8.
3
Solid phase in vitro mutagenesis using plasmid DNA template.使用质粒DNA模板的固相体外诱变
Nucleic Acids Res. 1990 Sep 11;18(17):5107-12. doi: 10.1093/nar/18.17.5107.
4
Site-directed mutagenesis of virtually any plasmid by eliminating a unique site.通过消除一个独特位点对几乎任何质粒进行定点诱变。
Anal Biochem. 1992 Jan;200(1):81-8. doi: 10.1016/0003-2697(92)90280-k.
5
Use of lambda exonuclease for efficient oligonucleotide-mediated site-directed deletion and point mutation of double-stranded DNA.使用λ核酸外切酶进行高效的寡核苷酸介导的双链DNA定点缺失和点突变。
DNA. 1987 Jun;6(3):273-9. doi: 10.1089/dna.1987.6.273.
6
Simple and efficient oligonucleotide-directed mutagenesis using one primer and circular plasmid DNA template.使用一个引物和环状质粒DNA模板进行简单高效的寡核苷酸定向诱变。
Gene Anal Tech. 1989 Jul-Aug;6(4):67-70. doi: 10.1016/0735-0651(89)90017-4.
7
Simple approach for oligonucleotide-directed mutagenesis of any double-stranded circular DNA.用于任何双链环状DNA寡核苷酸定向诱变的简易方法。
FEBS Lett. 1991 Jun 3;283(2):227-9. doi: 10.1016/0014-5793(91)80594-s.
8
Uracil-USE, an improved method for site-directed mutagenesis on double-stranded plasmid DNA.尿嘧啶使用法（Uracil-USE），一种用于双链质粒DNA定点诱变的改进方法。
Biotechniques. 1995 Mar;18(3):370-2.
9
A simple and efficient method for the oligodeoxyribonucleotide-directed mutagenesis of double-stranded plasmid DNA.一种用于双链质粒DNA的寡脱氧核糖核苷酸定向诱变的简单高效方法。
Gene. 1992 Nov 2;121(1):17-24. doi: 10.1016/0378-1119(92)90157-k.
10
Oligonucleotide-directed double-strand break repair in plasmids of Escherichia coli: a method for site-specific mutagenesis.大肠杆菌质粒中寡核苷酸定向双链断裂修复：一种位点特异性诱变方法。
Proc Natl Acad Sci U S A. 1986 Oct;83(19):7177-81. doi: 10.1073/pnas.83.19.7177.
引用本文的文献
1
DNA assembly using common laboratory bacteria: A re-emerging tool to simplify molecular cloning.利用常见的实验室细菌进行 DNA 组装：一种简化分子克隆的新兴工具。
J Biol Chem. 2019 Oct 18;294(42):15271-15281. doi: 10.1074/jbc.REV119.009109. Epub 2019 Sep 14.
2
A simple method for site-directed mutagenesis with double-stranded plasmid DNA.
Mol Biotechnol. 1998 Jun;9(3):235-41. doi: 10.1007/BF02915796.
本文引用的文献
1
Oligonucleotide-directed mutagenesis using M13-derived vectors: an efficient and general procedure for the production of point mutations in any fragment of DNA.使用M13衍生载体的寡核苷酸定向诱变：在任何DNA片段中产生点突变的高效通用方法。
Nucleic Acids Res. 1982 Oct 25;10(20):6487-500. doi: 10.1093/nar/10.20.6487.
2
Different base/base mismatches are corrected with different efficiencies by the methyl-directed DNA mismatch-repair system of E. coli.大肠杆菌的甲基化导向DNA错配修复系统对不同的碱基/碱基错配进行校正的效率不同。
Cell. 1984 Oct;38(3):879-87. doi: 10.1016/0092-8674(84)90283-6.
3
A simple and efficient procedure for saturation mutagenesis using mixed oligodeoxynucleotides.一种使用混合寡脱氧核苷酸进行饱和诱变的简单高效方法。
Gene. 1986;46(2-3):145-52. doi: 10.1016/0378-1119(86)90398-7.
4
Procedures for in vitro DNA mutagenesis of human leukocyte interferon sequences.人白细胞干扰素序列的体外DNA诱变程序。
Methods Enzymol. 1986;119:403-15. doi: 10.1016/0076-6879(86)19060-4.
5
A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions.DNA片段的体外制备及特异性诱变的通用方法：蛋白质与DNA相互作用的研究
Nucleic Acids Res. 1988 Aug 11;16(15):7351-67. doi: 10.1093/nar/16.15.7351.
6
DNA mismatch-repair in Escherichia coli counteracting the hydrolytic deamination of 5-methyl-cytosine residues.大肠杆菌中的DNA错配修复可对抗5-甲基胞嘧啶残基的水解脱氨作用。
EMBO J. 1987 Jun;6(6):1809-15. doi: 10.1002/j.1460-2075.1987.tb02435.x.
7
Construction of mutant and chimeric genes using the polymerase chain reaction.利用聚合酶链反应构建突变基因和嵌合基因。
Nucleic Acids Res. 1989 Jan 25;17(2):723-33. doi: 10.1093/nar/17.2.723.
8
Creation of phosphorylation sites in proteins: construction of a phosphorylatable human interferon alpha.
Proc Natl Acad Sci U S A. 1989 Jan;86(2):558-62. doi: 10.1073/pnas.86.2.558.
9
Analysis of any point mutation in DNA. The amplification refractory mutation system (ARMS).DNA中任何点突变的分析。扩增阻滞突变系统（ARMS）。
Nucleic Acids Res. 1989 Apr 11;17(7):2503-16. doi: 10.1093/nar/17.7.2503.
10
Site-directed mutagenesis by overlap extension using the polymerase chain reaction.利用聚合酶链反应通过重叠延伸进行定点诱变。
Gene. 1989 Apr 15;77(1):51-9. doi: 10.1016/0378-1119(89)90358-2.
Zotero 插件