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Rapid typing of tandemly repeated hypervariable loci by the polymerase chain reaction: application to the apolipoprotein B 3' hypervariable region.

Abstract：

:The 3' flanking region of the apolipoprotein B (apoB) gene contains a hypervariable region consisting of a variable number of tandemly repeated short A + T-rich DNA sequences (VNTRs). We present a general method that utilizes the polymerase chain reaction to rapidly and accurately type this and other VNTR loci. We use tailored oligonucleotides and thermostable Taq polymerase to amplify the targeted region. The amplification products are directly visualized after agarose gel electrophoresis. Twelve alleles were readily identified in a sample of 125 unrelated individuals. The alleles differ with respect to the length of the amplified gene region. This genetic variability is inherited in an autosomal codominant manner. DNA sequence data indicate that individual alleles differ in the number of repeat units and the sensitivity of the technique is such that alleles differing in length by only 32 base pairs are readily distinguishable. A system of nomenclature based on the number of repeat units is suggested; an allele containing 37 repeat units is designated 3' beta 37, one containing 35 units is 3' beta 35, and so on. The frequency distribution of the 12 apoB VNTR alleles is bimodal with peaks at 37 and 47 repeat units and a nadir near 43 repeat units. We estimate that the 3' apoB VNTR locus has a heterozygosity index of 0.75 and a polymorphic information content of 0.73. It is a highly informative marker for genetic linkage studies on chromosome 2 and clinical and epidemiological studies involving the apoB gene. The high sensitivity and inexpensive nature of this technique make it superior to traditional Southern blot analysis for typing the 3' apoB VNTR. The method described is also directly applicable for rapid typing of other VNTRs in the human genome.

journal_name

Proc Natl Acad Sci U S A

journal_title

Proceedings of the National Academy of Sciences of the United States of America

authors

Boerwinkle E,Xiong WJ,Fourest E,Chan L

doi

10.1073/pnas.86.1.212

subject

Has Abstract

pub_date

1989-01-01 00:00:00

pages

212-6

issue

eissn

0027-8424

issn

1091-6490

journal_volume

pub_type

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Rapid typing of tandemly repeated hypervariable loci by the polymerase chain reaction: application to the apolipoprotein B 3' hypervariable region.