镜像方向选择（MOS）：一种从抑制性消减杂交产生的文库中消除假阳性克隆的方法。
Mirror orientation selection (MOS): a method for eliminating false positive clones from libraries generated by suppression subtractive hybridization.
作者信息
Rebrikov D V, Britanova O V, Gurskaya N G, Lukyanov K A, Tarabykin V S, Lukyanov S A
机构信息
Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry RAS, Miklukho-Maklaya 16/10, 117871 Moscow, Russia.
出版信息
Nucleic Acids Res. 2000 Oct 15;28(20):E90. doi: 10.1093/nar/28.20.e90.
Abstract
Suppression subtractive hybridization (SSH) is one of the most powerful and popular methods for isolating differentially expressed transcripts. However, SSH-generated libraries typically contain some background clones representing non-differentially expressed transcripts. To overcome this problem we developed a simple procedure that substantially decreases the number of background clones. This method is based on the following difference between target and background cDNAs: each kind of background molecule has only one orientation with respect to the two different flanking adapter sequences used in SSH, while truly differentially expressed target cDNA fragments are represented by both sequence orientations. The described method selects the molecules that arose due to hybridization of such mirror-orientated molecules. The efficiency of this method was demonstrated in both model and real experimental subtractions.
摘要
抑制性消减杂交（SSH）是分离差异表达转录本最强大且最常用的方法之一。然而，SSH产生的文库通常包含一些代表非差异表达转录本的背景克隆。为克服这一问题，我们开发了一种简单的程序，可大幅减少背景克隆的数量。该方法基于目标cDNA与背景cDNA之间的以下差异：每种背景分子相对于SSH中使用的两种不同侧翼接头序列仅有一种方向，而真正差异表达的目标cDNA片段由两种序列方向代表。所描述的方法选择由这种镜像方向分子杂交产生的分子。该方法的效率在模型和实际实验消减中均得到了证明。
相似文献
1
Mirror orientation selection (MOS): a method for eliminating false positive clones from libraries generated by suppression subtractive hybridization.镜像方向选择（MOS）：一种从抑制性消减杂交产生的文库中消除假阳性克隆的方法。
Nucleic Acids Res. 2000 Oct 15;28(20):E90. doi: 10.1093/nar/28.20.e90.
2
Identification of Differential Genes by Suppression Subtractive Hybridization: IV. Mirror Orientation Selection (MOS).通过抑制性消减杂交技术鉴定差异基因：IV. 镜像定向选择（MOS）。
CSH Protoc. 2008 Jul 1;2008:pdb.prot4858. doi: 10.1101/pdb.prot4858.
3
Suppression subtractive hybridization: a versatile method for identifying differentially expressed genes.抑制性消减杂交：一种用于鉴定差异表达基因的通用方法。
Methods Enzymol. 1999;303:349-80. doi: 10.1016/s0076-6879(99)03022-0.
4
Subtractive cloning: new genes for studying inflammatory disorders.消减克隆：用于研究炎症性疾病的新基因
Ann Periodontol. 2002 Dec;7(1):17-28. doi: 10.1902/annals.2002.7.1.17.
5
Suppression subtractive hybridization.抑制性消减杂交
Methods Mol Biol. 2004;258:107-34. doi: 10.1385/1-59259-751-3:107.
6
Size-selection of cDNA libraries for the cloning of cDNAs after suppression subtractive hybridization.抑制消减杂交后用于克隆cDNA的cDNA文库的大小选择。
Biotechniques. 2003 Jul;35(1):72-8. doi: 10.2144/03351st02.
7
[Construction of suppression subtractive hybridization cDNA library of half-blood males of Dermacentor silvarum and analysis of differentially expressed genes].[森林革蜱半血雄蜱抑制性消减杂交cDNA文库的构建及差异表达基因分析]
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2014 Aug;32(4):274-9.
8
SSH adequacy to preimplantation mammalian development: scarce specific transcripts cloning despite irregular normalisation.SSH对哺乳动物植入前发育的充分性：尽管标准化不规律，但仍缺乏特异性转录本克隆。
BMC Genomics. 2005 Nov 8;6:155. doi: 10.1186/1471-2164-6-155.
9
Microarray hybridization with fractionated cDNA: enhanced identification of differentially expressed genes.用分级cDNA进行微阵列杂交：增强对差异表达基因的鉴定
Anal Biochem. 2000 Dec 1;287(1):32-7. doi: 10.1006/abio.2000.4831.
10
Construction of primary and subtracted cDNA libraries from early embryos.从早期胚胎构建初级和扣除cDNA文库。
Methods Enzymol. 1993;225:587-610. doi: 10.1016/0076-6879(93)25038-4.
引用本文的文献
1
Identification of genes associated with shell color in the black-lipped pearl oyster, Pinctada margaritifera.黑唇珠母贝（Pinctada margaritifera）中与壳色相关基因的鉴定。
BMC Genomics. 2015 Aug 1;16(1):568. doi: 10.1186/s12864-015-1776-x.
2
Subtractive and differential hybridization molecular analyses of Ceratitis capitata XX/XY versus XX embryos to search for male-specific early transcribed genes.对地中海实蝇XX/XY与XX胚胎进行消减杂交和差异杂交分子分析，以寻找雄性特异性早期转录基因。
BMC Genet. 2014;15 Suppl 2(Suppl 2):S5. doi: 10.1186/1471-2156-15-S2-S5. Epub 2014 Dec 1.
3
Development of a strain-specific real-time PCR assay for enumeration of a probiotic Lactobacillus reuteri in chicken feed and intestine.用于定量鸡饲料和肠道中益生菌罗伊氏乳杆菌的菌株特异性实时荧光定量PCR检测方法的建立。
PLoS One. 2014 Feb 27;9(2):e90208. doi: 10.1371/journal.pone.0090208. eCollection 2014.
4
Removing PCR for the elimination of undesired DNA fragments cycle by cycle.通过循环去除 PCR 以消除不需要的 DNA 片段。
Sci Rep. 2013;3:2303. doi: 10.1038/srep02303.
5
Methanol may function as a cross-kingdom signal.甲醇可能作为一种跨领域信号发挥作用。
PLoS One. 2012;7(4):e36122. doi: 10.1371/journal.pone.0036122. Epub 2012 Apr 26.
6
Enchytraeus albidus microarray: enrichment, design, annotation and database (EnchyBASE).白纹蚓 microarray：富集、设计、注释和数据库 (EnchyBASE)。
PLoS One. 2012;7(4):e34266. doi: 10.1371/journal.pone.0034266. Epub 2012 Apr 27.
7
Isolation and analysis of genes mainly expressed in adult mouse heart using subtractive hybridization cDNA library.利用消减杂交 cDNA 文库分离和分析主要在成年鼠心脏中表达的基因。
Mol Biol Rep. 2012 Aug;39(8):8065-74. doi: 10.1007/s11033-012-1653-5. Epub 2012 Apr 29.
8
Identification and expression analysis of methyl jasmonate responsive ESTs in paclitaxel producing Taxus cuspidata suspension culture cells.鉴定和茉莉酸甲酯响应 ESTs 在紫杉醇生产红豆杉悬浮培养细胞中的表达分析。
BMC Genomics. 2012 Apr 24;13:148. doi: 10.1186/1471-2164-13-148.
9
Airborne signals from a wounded leaf facilitate viral spreading and induce antibacterial resistance in neighboring plants.受伤叶片释放的空气信号可促进病毒的传播，并诱导邻近植物产生抗细菌的能力。
PLoS Pathog. 2012;8(4):e1002640. doi: 10.1371/journal.ppat.1002640. Epub 2012 Apr 5.
10
Influence of Wolbachia on host gene expression in an obligatory symbiosis.沃尔巴克氏体对专性共生中宿主基因表达的影响。
BMC Microbiol. 2012 Jan 18;12 Suppl 1(Suppl 1):S7. doi: 10.1186/1471-2180-12-S1-S7.
本文引用的文献
1
Suppression subtractive hybridization: a versatile method for identifying differentially expressed genes.抑制性消减杂交：一种用于鉴定差异表达基因的通用方法。
Methods Enzymol. 1999;303:349-80. doi: 10.1016/s0076-6879(99)03022-0.
2
A molecular pathway revealing a genetic basis for human cardiac and craniofacial defects.一条揭示人类心脏和颅面缺陷遗传基础的分子途径。
Science. 1999 Feb 19;283(5405):1158-61. doi: 10.1126/science.283.5405.1158.
3
Differential screening of a subtracted cDNA library: a method to search for genes preferentially expressed in multiple tissues.消减cDNA文库的差异筛选：一种寻找在多种组织中优先表达基因的方法。
Biotechniques. 1997 Dec;23(6):1084-6. doi: 10.2144/97236st05.
4
A high throughput screening for rarely transcribed differentially expressed genes.对低转录差异表达基因的高通量筛选。
Nucleic Acids Res. 1997 Jul 1;25(13):2598-602. doi: 10.1093/nar/25.13.2598.
5
A G-protein-coupled receptor for leukotriene B4 that mediates chemotaxis.一种介导趋化作用的白三烯B4 G蛋白偶联受体。
Nature. 1997 Jun 5;387(6633):620-4. doi: 10.1038/42506.
6
Construction of cDNA libraries from small amounts of total RNA using the suppression PCR effect.利用抑制性PCR效应从少量总RNA构建cDNA文库。
Biochem Biophys Res Commun. 1997 Jan 13;230(2):285-8. doi: 10.1006/bbrc.1996.5948.
7
Equalizing cDNA subtraction based on selective suppression of polymerase chain reaction: cloning of Jurkat cell transcripts induced by phytohemaglutinin and phorbol 12-myristate 13-acetate.基于聚合酶链反应选择性抑制的均等化cDNA消减：克隆由植物血凝素和佛波酯12-肉豆蔻酸酯13-乙酸酯诱导的Jurkat细胞转录本
Anal Biochem. 1996 Aug 15;240(1):90-7. doi: 10.1006/abio.1996.0334.
8
Molecule by molecule PCR amplification of complex DNA mixtures for direct sequencing: an approach to in vitro cloning.用于直接测序的复杂DNA混合物的逐分子PCR扩增：一种体外克隆方法。
Nucleic Acids Res. 1996 Jun 1;24(11):2194-5. doi: 10.1093/nar/24.11.2194.
9
Suppression subtractive hybridization: a method for generating differentially regulated or tissue-specific cDNA probes and libraries.抑制性消减杂交：一种用于生成差异调控或组织特异性cDNA探针及文库的方法。
Proc Natl Acad Sci U S A. 1996 Jun 11;93(12):6025-30. doi: 10.1073/pnas.93.12.6025.
10
An improved PCR method for walking in uncloned genomic DNA.一种用于在未克隆基因组DNA中进行步移的改进PCR方法。
Nucleic Acids Res. 1995 Mar 25;23(6):1087-8. doi: 10.1093/nar/23.6.1087.
Zotero 插件