一种简单高效的DNA化学诱变方法。
A simple and efficient method for chemical mutagenesis of DNA.
作者信息
Kadonaga J T, Knowles J R
出版信息
Nucleic Acids Res. 1985 Mar 11;13(5):1733-45. doi: 10.1093/nar/13.5.1733.
Abstract
A simple and efficient procedure for the generation of random GC to AT transition mutations in a specific DNA segment is described. A restriction fragment is inserted in each orientation into an M13 vector, single-stranded virion DNA from each recombinant phage is treated with methoxylamine, and, after reannealing of the mutagenized strands, a double-stranded restriction fragment is obtained. This methoxylamine-derivatized DNA segment is then joined with linearized M13 RF DNA, competent E. coli is transfected, and mutations are directly identified by sequencing of the phage DNA. Using this technique, single and double nucleotide substitutions were generated at a frequency greater than 50% in a 56-base pair segment of the signal codons of the TEM beta-lactamase.
摘要
本文描述了一种在特定DNA片段中产生随机GC到AT转换突变的简单高效方法。将一个限制性片段以每种方向插入到M13载体中，来自每个重组噬菌体的单链病毒体DNA用甲氧基胺处理，并且在诱变链重新退火后，获得双链限制性片段。然后将这个甲氧基胺衍生化的DNA片段与线性化的M13 RF DNA连接，转染感受态大肠杆菌，并通过对噬菌体DNA进行测序直接鉴定突变。使用该技术，在TEMβ-内酰胺酶信号密码子的56个碱基对片段中，单核苷酸和双核苷酸取代的产生频率大于50%。
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本文引用的文献
1
The chemical and mutagenic specificity of hydroxylamine.羟胺的化学特异性和诱变特异性。
Proc Natl Acad Sci U S A. 1961 Jun 15;47(6):845-55. doi: 10.1073/pnas.47.6.845.
2
Gap misrepair mutagenesis: efficient site-directed induction of transition, transversion, and frameshift mutations in vitro.缺口错配诱变：体外高效定点诱导转换、颠换和移码突变
Proc Natl Acad Sci U S A. 1982 Mar;79(5):1588-92. doi: 10.1073/pnas.79.5.1588.
3
Segment-specific mutagenesis: extensive mutagenesis of a lac promoter/operator element.片段特异性诱变：lac启动子/操纵元件的广泛诱变
Proc Natl Acad Sci U S A. 1982 Mar;79(5):1408-12. doi: 10.1073/pnas.79.5.1408.
4
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.
5
The role of the beta-lactamase signal sequence in the secretion of proteins by Escherichia coli.β-内酰胺酶信号序列在大肠杆菌蛋白质分泌中的作用。
J Biol Chem. 1984 Feb 25;259(4):2149-54.
6
A rapid and efficient method for region- and strand-specific mutagenesis of cloned DNA.一种用于克隆DNA区域特异性和链特异性诱变的快速高效方法。
EMBO J. 1982;1(4):433-7. doi: 10.1002/j.1460-2075.1982.tb01187.x.
7
New M13 vectors for cloning.用于克隆的新型M13载体。
Methods Enzymol. 1983;101:20-78. doi: 10.1016/0076-6879(83)01005-8.
8
Rapid and efficient cosmid cloning.快速高效的黏粒克隆
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9
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10
Rapid bacteriophage sedimentation in the presence of polyethylene glycol and its application to large-scale virus purification.聚乙二醇存在下噬菌体的快速沉降及其在大规模病毒纯化中的应用
Virology. 1970 Mar;40(3):734-44. doi: 10.1016/0042-6822(70)90218-7.
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