• 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

相似文献

1
An improved PCR-mutagenesis strategy for two-site mutagenesis or sequence swapping between related genes.一种用于双位点诱变或相关基因间序列交换的改进型聚合酶链式反应诱变策略。
Nucleic Acids Res. 1998 Apr 1;26(7):1848-50. doi: 10.1093/nar/26.7.1848.
2
Mutagenesis and gene fusion by megaprimer PCR.通过大引物PCR进行诱变和基因融合。
Methods Mol Biol. 1997;67:173-82. doi: 10.1385/0-89603-483-6:173.
3
Two-stage PCR protocol allowing introduction of multiple mutations, deletions and insertions using QuikChange Site-Directed Mutagenesis.采用定点突变技术引入多个突变、缺失和插入的两步PCR方案。
Biotechniques. 1999 Apr;26(4):680-2. doi: 10.2144/99264st03.
4
Combination primer polymerase chain reaction for multi-site mutagenesis of close proximity sites.用于近距离位点多位点诱变的组合引物聚合酶链反应
J Biomol Tech. 2005 Dec;16(4):336-40.
5
Site-directed mutagenesis by double polymerase chain reaction.通过双重聚合酶链反应进行的定点诱变
Mol Biotechnol. 1995 Feb;3(1):1-7. doi: 10.1007/BF02821329.
6
Polymerase chain reaction-based point mutagenesis protocol.基于聚合酶链反应的点突变实验方案。
Methods Enzymol. 1993;217:218-27. doi: 10.1016/0076-6879(93)17064-c.
7
Improvement of PCR reaction conditions for site-directed mutagenesis of big plasmids.提高大质粒定点突变 PCR 反应条件。
J Zhejiang Univ Sci B. 2012 Apr;13(4):244-7. doi: 10.1631/jzus.B1100180.
8
An improved PCR-based method for site directed mutagenesis using megaprimers.一种改进的基于聚合酶链式反应（PCR）的使用大引物进行定点诱变的方法。
Mol Cell Probes. 1998 Dec;12(6):345-8. doi: 10.1006/mcpr.1998.0187.
9
In vitro scanning-saturation mutagenesis.体外扫描饱和诱变
Methods Mol Biol. 2002;182:149-63. doi: 10.1385/1-59259-194-9:149.
10
A novel megaprimed and ligase-free, PCR-based, site-directed mutagenesis method.一种新型的基于PCR的、无需连接酶的、采用大引物法的定点诱变方法。
Anal Biochem. 2008 Apr 15;375(2):376-8. doi: 10.1016/j.ab.2007.12.013. Epub 2007 Dec 23.

引用本文的文献

1
Influence of mutations at different distances from the active center on the activity and stability of laccase 13B22.活性中心不同距离处的突变对漆酶13B22活性和稳定性的影响。
Bioresour Bioprocess. 2025 May 27;12(1):47. doi: 10.1186/s40643-025-00893-6.
2
Enhancing the endo-activity of the thermophilic chitinase to yield chitooligosaccharides with high degrees of polymerization.增强嗜热几丁质酶的内切活性以产生具有高聚合度的壳寡糖。
Bioresour Bioprocess. 2024 Mar 7;11(1):29. doi: 10.1186/s40643-024-00735-x.
3
Biochemical and cellular insights into the Baz2B protein, a non-catalytic subunit of the chromatin remodeling complex.Baz2B 蛋白的生化和细胞内研究，该蛋白是非染色质重塑复合物的非催化亚基。
Nucleic Acids Res. 2024 Jan 11;52(1):337-354. doi: 10.1093/nar/gkad1096.
4
MPEPE, a predictive approach to improve protein expression in based on deep learning.MPEPE，一种基于深度学习提高蛋白质表达的预测方法。 （你提供的原文中“in based on”表述有误，推测可能是“in vitro”之类的，这里按照纠正后的意思翻译）
Comput Struct Biotechnol J. 2022 Mar 1;20:1142-1153. doi: 10.1016/j.csbj.2022.02.030. eCollection 2022.
5
The nucleotide messenger (p)ppGpp is an anti-inducer of the purine synthesis transcription regulator PurR in Bacillus.核苷酸信使 (p)ppGpp 是芽孢杆菌嘌呤合成转录调控因子 PurR 的抗诱导物。
Nucleic Acids Res. 2022 Jan 25;50(2):847-866. doi: 10.1093/nar/gkab1281.
6
uPIC-M: Efficient and Scalable Preparation of Clonal Single Mutant Libraries for High-Throughput Protein Biochemistry.uPIC-M：用于高通量蛋白质生物化学的克隆单突变体文库的高效且可扩展制备
ACS Omega. 2021 Nov 2;6(45):30542-30554. doi: 10.1021/acsomega.1c04180. eCollection 2021 Nov 16.
7
Redox controls RecA protein activity via reversible oxidation of its methionine residues.氧化还原通过其蛋氨酸残基的可逆氧化来控制 RecA 蛋白的活性。
Elife. 2021 Feb 19;10:e63747. doi: 10.7554/eLife.63747.
8
Uncoupling sodium channel dimers restores the phenotype of a pain-linked Na 1.7 channel mutation.去偶联钠离子通道二聚体可恢复与疼痛相关的 Na 1.7 通道突变的表型。
Br J Pharmacol. 2020 Oct;177(19):4481-4496. doi: 10.1111/bph.15196. Epub 2020 Aug 24.
9
Molecular Mechanism of Regulation of the Purine Salvage Enzyme XPRT by the Alarmones pppGpp, ppGpp, and pGpp.嘌呤补救酶 XPRT 的调节的分子机制由警报素 pppGpp、ppGpp 和 pGpp 调控。
J Mol Biol. 2020 Jun 26;432(14):4108-4126. doi: 10.1016/j.jmb.2020.05.013. Epub 2020 May 21.
10
Bridge helix arginines play a critical role in Cas9 sensitivity to mismatches.桥螺旋精氨酸在 Cas9 对错配的敏感性中起着关键作用。
Nat Chem Biol. 2020 May;16(5):587-595. doi: 10.1038/s41589-020-0490-4. Epub 2020 Mar 2.

本文引用的文献

1
Random-priming in vitro recombination: an effective tool for directed evolution.随机引物体外重组：定向进化的有效工具。
Nucleic Acids Res. 1998 Jan 15;26(2):681-3. doi: 10.1093/nar/26.2.681.
2
Modified base compositions at degenerate positions of a mutagenic oligonucleotide enhance randomness in site-saturation mutagenesis.诱变寡核苷酸简并位点处的碱基组成修饰可增强位点饱和诱变的随机性。
Nucleic Acids Res. 1998 Jan 15;26(2):576-81. doi: 10.1093/nar/26.2.576.
3
Rapid and efficient site-directed mutagenesis by single-tube 'megaprimer' PCR method.通过单管“大引物”PCR 方法进行快速高效的定点诱变
Nucleic Acids Res. 1997 Aug 15;25(16):3371-2. doi: 10.1093/nar/25.16.3371.
4
PCR mutagenesis: treatment of the megaprimer with mung bean nuclease improves yield.聚合酶链反应诱变：用绿豆核酸酶处理大引物可提高产量。
Biotechniques. 1997 Jun;22(6):1054-6. doi: 10.2144/97226bm09.
5
A rapid and efficient method for site-directed mutagenesis using one-step overlap extension PCR.一种使用一步重叠延伸PCR进行定点诱变的快速高效方法。
Nucleic Acids Res. 1997 Jun 1;25(11):2227-8. doi: 10.1093/nar/25.11.2227.
6
"Megaprimer" method of PCR-based mutagenesis: the concentration of megaprimer is a critical factor.基于聚合酶链反应（PCR）的诱变“大引物”方法：大引物的浓度是一个关键因素。
Biotechniques. 1997 Mar;22(3):438, 442. doi: 10.2144/97223bm13.
7
Genes in two MHC class I regions control recognition of a single rat NK cell allodeterminant.两个MHC I类区域中的基因控制对单个大鼠自然杀伤细胞同种异体决定簇的识别。
Int Immunol. 1996 Nov;8(11):1779-85. doi: 10.1093/intimm/8.11.1779.
8
An efficient PCR mutagenesis strategy without gel purification [correction of "purificiation"] step that is amenable to automation.一种无需凝胶纯化步骤且适用于自动化的高效PCR诱变策略。 [“purificiation”的更正为“purification”]
Nucleic Acids Res. 1996 Aug 15;24(16):3276-7. doi: 10.1093/nar/24.16.3276.
9
The rat MHC haplotype RT1c expresses two classical class I molecules.大鼠主要组织相容性复合体单倍型RT1c表达两种经典的I类分子。
J Immunol. 1996 Aug 15;157(4):1551-8.
10
Two versatile eukaryotic vectors permitting epitope tagging, radiolabelling and nuclear localisation of expressed proteins.两种通用的真核载体，可实现对表达蛋白进行表位标记、放射性标记和核定位。
Gene. 1996 Feb 12;168(2):165-7. doi: 10.1016/0378-1119(95)00764-4.

一种用于双位点诱变或相关基因间序列交换的改进型聚合酶链式反应诱变策略。

An improved PCR-mutagenesis strategy for two-site mutagenesis or sequence swapping between related genes.

作者信息

Kirsch R D, Joly E

机构信息

The Babraham Institute, Babraham, Cambridge CB2 4AT, UK.

出版信息

Nucleic Acids Res. 1998 Apr 1;26(7):1848-50. doi: 10.1093/nar/26.7.1848.

DOI:10.1093/nar/26.7.1848
PMID:9512562
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC147442/
Abstract

The QuikChangeTM protocol is one of the simplest and fastest methods for site-directed mutagenesis, but introduces mutations at only one site at a time, and requires two HPLC-purified complementary oligonucleotides. Here, we describe that this method can be used with non-overlapping oligonucleotides. By doing this, two separate sites can be mutagenised simultaneously, or money can be saved by using a second 'standard' oligonucleotide. By a further modification, we have also used the QuikChangeTM approach to exchange DNA sequences between closely related genes.

摘要

快速定点突变（QuikChangeTM）协议是定点突变最简单、最快的方法之一，但每次仅在一个位点引入突变，且需要两条经高效液相色谱（HPLC）纯化的互补寡核苷酸。在此，我们描述了该方法可用于非重叠寡核苷酸。通过这样做，可以同时诱变两个独立的位点，或者通过使用第二条“标准”寡核苷酸节省成本。通过进一步改进，我们还采用了快速定点突变（QuikChangeTM）方法在密切相关的基因之间交换DNA序列。