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通过聚合酶链反应和毛细管电泳对人载脂蛋白B基因中可变数目串联重复序列位点进行快速分型用于心脏病的DNA诊断

Rapid typing of variable number of tandem repeat locus in the human apolipoprotein B gene for DNA diagnosis of heart disease by polymerase chain reaction and capillary electrophoresis.

作者信息

Baba Y, Tomisaki R, Sumita C, Morimoto I, Sugita S, Tsuhako M, Miki T, Ogihara T

机构信息

Department of Chemistry, Kobe Pharmaceutical University, Japan.

出版信息

Electrophoresis. 1995 Aug;16(8):1437-40. doi: 10.1002/elps.11501601237.

Abstract

The apolipoprotein B (apoB) variable number of tandem repeat (VNTR) alleles containing larger repeat units is a risk factor for coronary heart disease. Capillary electrophoresis (CE) in entangled polymer solution was applied to the analysis of polymerase chain reaction (PCR) amplified apoB VNTR locus for DNA diagnosis of heart disease. The CE separation gives an excellent resolution of two alleles differing by one or two 16 bp repeat units in the DNA size range up to 600 bp with high speed. The apoB alleles differing in length by 2 or 4 repeat units are readily distinguishable by CE in the DNA size range from 600 to 1000 bp. The plate number achieved was 1 million plates per meter. CE combining with PCR provides an excellent technique for accurate determination of the number of repeat units of apoB VNTR alleles and differentiation of heterozygous from homozygous individuals. Using the CE technique, the apoB VNTR loci from some individuals in genotyping were examined towards precise DNA diagnosis for coronary heart disease.

摘要

载脂蛋白B(apoB)串联重复序列(VNTR)中包含较大重复单元的等位基因是冠心病的一个风险因素。采用在缠结聚合物溶液中的毛细管电泳(CE)对聚合酶链反应(PCR)扩增的apoB VNTR基因座进行分析,用于心脏病的DNA诊断。CE分离能够在高达600 bp的DNA大小范围内,以高速出色地分辨出相差一个或两个16 bp重复单元的两个等位基因。在600至1000 bp的DNA大小范围内,相差2个或4个重复单元的apoB等位基因很容易通过CE区分开来。实现的板数为每米100万块板。CE与PCR相结合为准确确定apoB VNTR等位基因的重复单元数量以及区分杂合子和纯合子个体提供了一项出色的技术。利用CE技术,对一些个体基因分型中的apoB VNTR基因座进行了检测,以用于冠心病的精确DNA诊断。

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