反向聚合酶链反应的遗传学应用
Genetic applications of an inverse polymerase chain reaction.
作者信息
Ochman H, Gerber A S, Hartl D L
机构信息
Department of Genetics, Washington University School of Medicine, St. Louis, Missouri 63110.
出版信息
Genetics. 1988 Nov;120(3):621-3. doi: 10.1093/genetics/120.3.621.
Abstract
A method is presented for the rapid in vitro amplification of DNA sequences that flank a region of known sequence. The method uses the polymerase chain reaction (PCR), but it has the primers oriented in the reverse direction of the usual orientation. The template for the reverse primers is a restriction fragment that has been ligated upon itself to form a circle. This procedure of inverse PCR (IPCR) has many applications in molecular genetics, for example, the amplification and identification of sequences flanking transposable elements. In this paper we show the feasibility of IPCR by amplifying the sequences that flank an IS1 element in the genome of a natural isolate of Escherichia coli.
摘要
本文介绍了一种用于快速体外扩增已知序列区域侧翼DNA序列的方法。该方法使用聚合酶链反应（PCR），但其引物的方向与通常的方向相反。反向引物的模板是一个已自身连接形成环状的限制性片段。这种反向PCR（IPCR）方法在分子遗传学中有许多应用，例如，扩增和鉴定转座元件侧翼的序列。在本文中，我们通过扩增大肠杆菌自然分离株基因组中IS1元件侧翼的序列，展示了IPCR的可行性。
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本文引用的文献
1
Directional cloning of DNA fragments at a large distance from an initial probe: a circularization method.从初始探针远距离定向克隆DNA片段：一种环化方法。
Proc Natl Acad Sci U S A. 1984 Nov;81(21):6812-6. doi: 10.1073/pnas.81.21.6812.
2
Direct cloning and sequence analysis of enzymatically amplified genomic sequences.酶促扩增基因组序列的直接克隆与序列分析
Science. 1986 Sep 5;233(4768):1076-8. doi: 10.1126/science.3461561.
3
Specific synthesis of DNA in vitro via a polymerase-catalyzed chain reaction.通过聚合酶催化的链式反应在体外特异性合成DNA。
Methods Enzymol. 1987;155:335-50. doi: 10.1016/0076-6879(87)55023-6.
4
Distribution and abundance of insertion sequences among natural isolates of Escherichia coli.大肠杆菌自然分离株中插入序列的分布与丰度
Genetics. 1987 Jan;115(1):51-63. doi: 10.1093/genetics/115.1.51.
5
Enzymatic amplification of beta-globin genomic sequences and restriction site analysis for diagnosis of sickle cell anemia.用于镰状细胞贫血诊断的β-珠蛋白基因组序列的酶促扩增及限制性酶切位点分析。
Science. 1985 Dec 20;230(4732):1350-4. doi: 10.1126/science.2999980.
6
Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase.使用热稳定DNA聚合酶进行引物引导的DNA酶促扩增。
Science. 1988 Jan 29;239(4839):487-91. doi: 10.1126/science.2448875.
7
Nucleotide sequence of an insertion element, IS1.插入元件IS1的核苷酸序列。
Proc Natl Acad Sci U S A. 1978 Feb;75(2):615-9. doi: 10.1073/pnas.75.2.615.
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