Skip to main content
U.S. Department of Energy
Office of Scientific and Technical Information

Factors affecting mutational specificity in mammalian cells: (Informal technical progress report), February 1, 1986-January 31, 1987

Technical Report ·
OSTI ID:7024799
We analyzed the different c-H-ras mutations produced in cells after treatment with chemical carcinogens. The overall goal of this work is an understanding of the changes produced by environmental mutagens and carcinogens to learn how closely the results obtained with bacterial and cellular assay systems apply to the in vivo situation. Our work continues to utilize the technique for the study of DNA synthesis termination in vitro developed in this laboraory and utilized for the past several years. This technology can be characterized as a Sanger dideoxy sequencing technique with mutation induced lesions in the template strand serving as chain terminators instead of the dideoxynucleotides used in the Sanger technique to terminate the growing strand. We have added to this technique a modification for the study of termination on double stranded templates as possibly modeling more closely the natural situation. For this modification, double stranded M13 DNA is split once with a restriction enzyme which makes a unique cut and the linearized DNA is hybridized with single stranded circular DNA to make a uniquely nicked double stranded circle with a single 3'OH to serve as a primer for DNA synthesis which can occur by either strand displacement or nick translation.
Research Organization:
Chicago Univ., IL (USA)
DOE Contract Number:
AC02-76EV02040
OSTI ID:
7024799
Report Number(s):
DOE/EV/02040-T2; ON: DE88012387
Country of Publication:
United States
Language:
English