RAPID CLONING OF HLA-A,B CDNA BY USING THE POLYMERASE CHAIN-REACTION - FREQUENCY AND NATURE OF ERRORS PRODUCED IN AMPLIFICATION

被引：323

作者：

ENNIS, PD ^{[1
]}

ZEMMOUR, J ^{[1
]}

SALTER, RD ^{[1
]}

PARHAM, P ^{[1
]}

机构：

[1] STANFORD UNIV,DEPT CELL BIOL,STANFORD,CA 94305

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 1990年 / 87卷 / 07期

关键词：

gene families; histocompatibility; polymorphism; recombination;

D O I：

10.1073/pnas.87.7.2833

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

A method for cloning full-length HLA-A,B cDNA (1.1 kilobases) by using the polymerase chain reaction (PCR) is described. Six HLA-A,B alleles (HLA-A2, -A25, -B7, -B37, -B51, and -B57) were cloned, and their structures were determined. Multiple PCR clones for each allele were sequenced to obtain both an accurate consensus sequence and an 'authentic' clone having that sequence. Sequences from 50 clones encoding five different alleles permit assessment of the frequency and nature of PCR-produced errors. These include recombinations, deletions, and insertions in addition to point substitutions. Authentic clones were obtained at a frequency of between 30% and 70%, and analysis of three or four clones generally should be sufficient for characterization of an allele.

引用

页码：2833 / 2837

页数：5

共 27 条

[1] STRUCTURE OF THE HUMAN CLASS-I HISTOCOMPATIBILITY ANTIGEN, HLA-A2 [J].