Use of the polymerase chain reaction technique to create base-specific ras oncogene mutations.
Use of the polymerase chain reaction technique to create base-specific ras oncogene mutations.
复制标题
使用聚合酶链式反应技术创建碱基特异性 ras 癌基因突变。
DOI：
10.1089/dna.1.1988.7.515
复制
发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Benz,CC
中科院分区：
文献类型：
--
作者：
Rochlitz,CF;Scott,GK;Dodson,JM;Benz,CC
A modification of the polymerase chain reaction technique (PCR) technique, a primer-mediated enzymatic amplification of specific target sequences in genomic DNA, was used to introduce point mutations into copies of humanrasoncogene sequences, and to amplify these mutated copies approximately 106-fold. Of the two flanking oligomers used to amplify the DNA, one contained a single base mismatch with the targeted gene segment, either codon 12 or 61 from the Kirstenrasoncogene. Double-stranded fragments harboring any point mutation can be generated using the appropriate oligomer and constitute > 99.999% of the PCR-amplified fragments. The amplified DNA fragments are readily slot-blotted to nylon membranes to serve as positive hybridization controls in the search for gene mutations in human tissue specimens. The synthesis of single base mismatched DNA fragments was also used to demonstrate that oncogene mutations can be detected from mixed DNA populations as might be present in primary tumor specimens, even when the mutation of interest is present in only 5% of the amplified sample DNA.
