Bioenvironmental Hazards and DNA Repair
Abstract
Information is needed on mechanisms by which humans respond to exposure of their cellular genes to toxic chemicals in the environment. The principal DNA repair mechanism in organisms is excision repair and evidence has accumulated that these mechanisms are highly conserved. Using the fruit fly as a eukaryotic model, we undertook the molecular cloning of excision repair genes on the basis of their potential structural and functional similarity to the well- characterized excision repair genes in yeast. cDNA libraries were constructed from mRNA isolated from Drosophila embryos in a yeast expression vector, pYES 2. 0, and subsequently used to rescue a yeast rad3 mutant strain, known to be defective in excision repair. Although the equivalent of four genome equivalents were screened for complementation, no functional cognate Drosophila gene was recovered. On the basis of conserved DNA sequence between the yeast RAD3 and the human ERCC-2 genes, the polymerase chain reaction was used to recover a Drosophila cognate sequence. Using our CDNA library as template, a single NA band was identified. We interpret this to mean that Drosophila does have a RAD3 cognate but it was not represented in a functional form in our cDNA library to allow rescue of the rad3 mutant strain .
Document Details
- Document Type
- Technical Report
- Publication Date
- Apr 15, 1993
- Accession Number
- ADA264061
Entities
People
- P. D. Smith
Organizations
- Wayne State University