RAPID SCREENING OF MUTANT N-RAS ALLELES BY ANALYSIS OF PCR-INDUCED RESTRICTION SITES - ALLELE SPECIFIC RESTRICTION ANALYSIS (ASRA)

被引：18

作者：

TODD, AV

ILAND, HJ

机构：

[1] The Kanematsu Laboratories, Royal Prince Alfred Hospital, Camperdown

来源：

LEUKEMIA & LYMPHOMA | 1991年 / 3卷 / 04期

基金：

英国医学研究理事会;

关键词：

N-RAS; MUTATION; ACUTE MYELOID LEUKEMIA; POLYMERASE CHAIN REACTION; ALLELE SPECIFIC RESTRICTION ANALYSIS;

D O I：

10.3109/10428199109107917

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

We have developed a rapid screening method for analysis of codon 12, 13 and 61 N-ras gene mutations, since these mutations have been observed in approximately 25% of patients with acute myeloid leukemia and myelodysplastic syndromes. The method, termed allele specific restriction analysis (ASRA), involves polymerase chain reaction amplification of DNA or RNA using a mismatched primer which introduces appropriately positioned base substitutions in N-ras and creates a restriction site provided the adjacent sequence is normal. Simultaneous analysis of codons 12 and 61 is also possible by the use of a multiprimer reaction mixture. Resistance of the amplified product to digestion indicates the presence of a mutation in the original template. Since ASRA allows simultaneous analysis of mutant and wild type sequences in DNA and RNA, an estimate of the ratio of gene copies and relative expression of N-ras alleles can be obtained for heterozygous individuals. © 1991 Informa UK Ltd All rights reserved: reproduction in whole or part not permitted.

引用

页码：293 / 300

页数：8

共 17 条

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