Analysis of Salmonella typhimurium hisD3052 revertants: The use of oligodeoxyribonucleotide colony hybridization, PCR, and direct sequencing in mutational analysis

Environmental and Molecular Mutagenesis - Tập 18 Số 4 - Trang 224-230 - 1991
Eugene Kupchella1, Thomas A. Cebula2
1Molecular Biology Branch, Division of Microbiology, Center for Food Safety and Applied Nutrition, Food and Drug Administration, Washington, D.C.
2Molecular Biology Branch, HFF‐235, Division of Microbiology, Center for Food Safety and Applied Nutrition, Food and Drug Administration, 200 C Street, S.W., Washington, D.C. 20204

Tóm tắt

Abstract

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 oligodeoxy‐ribonucleotide 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.

Từ khóa


Tài liệu tham khảo

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Thông tin xuất bản

Environmental and Molecular Mutagenesis

Tập 18 Số 4

224-230

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