Kupchella E, Cebula T A
Molecular Biology Branch, Food and Drug Administration, Washington, D.C. 20204.
Environ Mol Mutagen. 1991;18(4):224-30. doi: 10.1002/em.2850180404.
A rapid method for determining the DNA sequences of Salmonella typhimurium hisD3052 revertants is presented. DNA colony hybridization was used to analyze revertants previously studied by Isono and Yourno [Proc Natl Acad Sci USA 71:1612-1617, 1974]. Synthetic oligodeoxyribonucleotide probes (18-mers) were able to distinguish sequences that differed by a single base pair. Mutant his sequences not identified by probing analysis were amplified using polymerase chain reaction (PCR) and directly sequenced. The combined use of DNA-colony hybridization and direct sequencing offers a precise and rapid means for the molecular characterization of hisD3052 revertants.
本文介绍了一种快速测定鼠伤寒沙门氏菌hisD3052回复突变体DNA序列的方法。采用DNA菌落杂交技术对Isono和Yourno先前研究过的回复突变体进行分析[《美国国家科学院院刊》71:1612 - 1617, 1974]。合成的寡脱氧核糖核苷酸探针（18聚体）能够区分相差一个碱基对的序列。未通过探针分析鉴定出的突变his序列，使用聚合酶链反应（PCR）进行扩增并直接测序。DNA菌落杂交和直接测序相结合的方法为hisD3052回复突变体的分子特征分析提供了一种精确且快速的手段。
