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。
Oligonucleotide-directed double-strand break repair in plasmids of Escherichia coli: a method for site-specific mutagenesis.
作者信息
Mandecki W
出版信息
Proc Natl Acad Sci U S A. 1986 Oct;83(19):7177-81. doi: 10.1073/pnas.83.19.7177.
Abstract
A DNA double-strand break can be efficiently repaired in Escherichia coli if an oligodeoxyribonucleotide is provided to direct the repair. The oligonucleotide must be at least 20 residues long and have a sequence identical to sequences flanking the break. The phenomenon can be used to introduce defined mutations into DNA in the area of a double-strand break. To obtain mutants, the oligonucleotide that carries a mutation and the denatured linearized plasmid DNA are introduced into E. coli by transformation. No enzymatic manipulation in vitro is required. The mutants can constitute up to 98% of the total number of transformants obtained. The efficiency of mutagenesis decreases as the distance between the mutation and the plasmid cleavage site increases. The universality of the method was tested by introducing mutations into four genes, using four plasmids and three E. coli strains, as well as eight restriction enzymes to linearize DNA. Several models of the oligonucleotide-directed DNA double-strand break repair are discussed.
摘要
如果提供一个寡脱氧核糖核苷酸来指导修复，DNA双链断裂在大肠杆菌中可以得到有效修复。该寡核苷酸必须至少20个残基长，并且具有与断裂侧翼序列相同的序列。这种现象可用于在双链断裂区域的DNA中引入特定突变。为了获得突变体，将携带突变的寡核苷酸和变性的线性化质粒DNA通过转化引入大肠杆菌。无需体外酶促操作。突变体可占所获得转化体总数的98%。随着突变与质粒切割位点之间距离的增加，诱变效率降低。通过使用四个质粒、三个大肠杆菌菌株以及八种限制酶线性化DNA，将突变引入四个基因，测试了该方法的通用性。讨论了几种寡核苷酸指导的DNA双链断裂修复模型。
相似文献
1
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.
2
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.
3
Oligonucleotide-Directed Mutagenesis by Elimination of a Unique Restriction Site (USE Mutagenesis).通过消除独特限制酶切位点进行寡核苷酸定向诱变（USE诱变）。
Cold Spring Harb Protoc. 2019 Jan 2;2019(1):2019/1/pdb.prot097782. doi: 10.1101/pdb.prot097782.
4
Repair of the plasmid pBR322 damaged by gamma-irradiation or by restriction endonucleases using different recombination-proficient E. coli strains.使用不同的重组缺陷型大肠杆菌菌株修复受γ射线或限制性内切酶损伤的质粒pBR322。
Mutat Res. 1988 Nov;194(3):193-205. doi: 10.1016/0167-8817(88)90021-1.
5
DNA gap repair in Escherichia coli for multiplex site-directed mutagenesis.大肠杆菌中用于多重位点定向诱变的 DNA 缺口修复。
FASEB J. 2020 May;34(5):6351-6368. doi: 10.1096/fj.201902260R. Epub 2020 Mar 13.
6
Double-strand breaks in plasmid DNA and the induction of deletions.质粒DNA中的双链断裂与缺失的诱导
Mutat Res. 1993 May;299(3-4):233-50. doi: 10.1016/0165-1218(93)90100-r.
7
Genetic applications of yeast transformation with linear and gapped plasmids.使用线性和缺口质粒进行酵母转化的遗传学应用。
Methods Enzymol. 1983;101:228-45. doi: 10.1016/0076-6879(83)01017-4.
8
Transformation of yeast with linearized plasmid DNA. Formation of inverted dimers and recombinant plasmid products.用线性化质粒DNA转化酵母。形成反向二聚体和重组质粒产物。
J Mol Biol. 1985 Aug 5;184(3):375-87. doi: 10.1016/0022-2836(85)90288-8.
9
Mutagenesis in Escherichia coli by three O6-substituted guanines in double-stranded or gapped plasmids.
Biochemistry. 1995 Jul 11;34(27):8924-30. doi: 10.1021/bi00027a045.
10
Deletion loop mutagenesis: a novel method for the construction of point mutations using deletion mutants.缺失环诱变：一种利用缺失突变体构建点突变的新方法。
Nucleic Acids Res. 1982 Sep 11;10(17):5161-71. doi: 10.1093/nar/10.17.5161.
引用本文的文献
1
On the Origins of Homology Directed Repair in Mammalian Cells.在哺乳动物细胞中同源定向修复的起源。
Int J Mol Sci. 2021 Mar 25;22(7):3348. doi: 10.3390/ijms22073348.
2
A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells.一种用于检测人细胞中由CRISPR/Cas9和单链寡脱氧核苷酸催化的点突变修复作用下的现场诱变性的标准方法。
J Vis Exp. 2017 Aug 25(126):56195. doi: 10.3791/56195.
3
Genetic spell-checking: gene editing using single-stranded DNA oligonucleotides.基因拼写检查：使用单链DNA寡核苷酸进行基因编辑。
Plant Biotechnol J. 2016 Feb;14(2):463-70. doi: 10.1111/pbi.12473. Epub 2015 Sep 24.
4
In vivo gene repair of point and frameshift mutations directed by chimeric RNA/DNA oligonucleotides and modified single-stranded oligonucleotides.由嵌合RNA/DNA寡核苷酸和修饰的单链寡核苷酸引导的点突变和移码突变的体内基因修复。
Nucleic Acids Res. 2001 Oct 15;29(20):4238-50. doi: 10.1093/nar/29.20.4238.
5
Mapping the multimerization domains of the Gag protein of yeast retrotransposon Ty1.绘制酵母逆转录转座子Ty1的Gag蛋白多聚化结构域图谱。
J Virol. 1997 Jan;71(1):812-7. doi: 10.1128/JVI.71.1.812-817.1997.
6
Protection of cats against feline leukemia virus by vaccination with a canarypox virus recombinant, ALVAC-FL.用金丝雀痘病毒重组体ALVAC-FL对猫进行疫苗接种以预防猫白血病病毒。
J Virol. 1993 Apr;67(4):2370-5. doi: 10.1128/JVI.67.4.2370-2375.1993.
7
A rare tRNA-Arg(CCU) that regulates Ty1 element ribosomal frameshifting is essential for Ty1 retrotransposition in Saccharomyces cerevisiae.一种调控Ty1元件核糖体移码的罕见tRNA-Arg(CCU)对酿酒酵母中的Ty1逆转录转座至关重要。
Genetics. 1993 Oct;135(2):309-20. doi: 10.1093/genetics/135.2.309.
8
Alterations of highly conserved residues in the regulatory domain of nitrogen regulator I (NtrC) of Escherichia coli.大肠杆菌氮调节蛋白I（NtrC）调节结构域中高度保守残基的改变。
J Bacteriol. 1993 May;175(9):2692-701. doi: 10.1128/jb.175.9.2692-2701.1993.
9
Escherichia coli gpt gene provides dominant selection for vaccinia virus open reading frame expression vectors.大肠杆菌gpt基因可为痘苗病毒开放阅读框表达载体提供显性选择。
J Virol. 1988 Jun;62(6):1849-54. doi: 10.1128/JVI.62.6.1849-1854.1988.
10
Alteration of the amino terminus of the mature sequence of a periplasmic protein can severely affect protein export in Escherichia coli.周质蛋白成熟序列氨基末端的改变会严重影响大肠杆菌中的蛋白质输出。
Proc Natl Acad Sci U S A. 1988 Oct;85(20):7685-9. doi: 10.1073/pnas.85.20.7685.
本文引用的文献
1
Analysis of gene control signals by DNA fusion and cloning in Escherichia coli.通过在大肠杆菌中进行DNA融合和克隆分析基因控制信号。
J Mol Biol. 1980 Apr;138(2):179-207. doi: 10.1016/0022-2836(80)90283-1.
2
DNA sequences from the str operon of Escherichia coli.来自大肠杆菌str操纵子的DNA序列。
J Biol Chem. 1980 May 25;255(10):4660-6.
3
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.
4
Recombination-dependent recircularization of linearized pBR322 plasmid DNA following transformation of Escherichia coli.线性化的pBR322质粒DNA在转化大肠杆菌后依赖重组的再环化
Mol Gen Genet. 1984;194(1-2):211-8. doi: 10.1007/BF00383519.
5
The pUC plasmids, an M13mp7-derived system for insertion mutagenesis and sequencing with synthetic universal primers.pUC质粒，一种源自M13mp7的用于插入诱变和使用合成通用引物进行测序的系统。
Gene. 1982 Oct;19(3):259-68. doi: 10.1016/0378-1119(82)90015-4.
6
Yeast transformation: a model system for the study of recombination.酵母转化：用于重组研究的模型系统。
Proc Natl Acad Sci U S A. 1981 Oct;78(10):6354-8. doi: 10.1073/pnas.78.10.6354.
7
A plasmid cloning vector for the direct selection of strains carrying recombinant plasmids.一种用于直接筛选携带重组质粒菌株的质粒克隆载体。
Gene. 1981 Oct;15(1):99-102. doi: 10.1016/0378-1119(81)90108-6.
8
Calcium-dependent bacteriophage DNA infection.钙依赖性噬菌体DNA感染。
J Mol Biol. 1970 Oct 14;53(1):159-62. doi: 10.1016/0022-2836(70)90051-3.
9
Supercoil sequencing: a fast and simple method for sequencing plasmid DNA.超螺旋测序：一种用于质粒DNA测序的快速简便方法。
DNA. 1985 Apr;4(2):165-70. doi: 10.1089/dna.1985.4.165.
10
Chemical synthesis of a gene encoding the human complement fragment C5a and its expression in Escherichia coli.
Proc Natl Acad Sci U S A. 1985 Jun;82(11):3543-7. doi: 10.1073/pnas.82.11.3543.
Zotero 插件