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M Nolden

Publications and source records attributed to M Nolden.

3 recordsLinked to original sources

Zero background yeast reporter plasmids.

UAS-less reporter plasmids are widespread and powerful tools for the identification and analysis of binding sites for transcriptional activators. The common reporter plasmids for the yeast Saccharomyces cerevisiae are multicopy (2mu) vectors with the CYC1 core promoter upstream of the lacZ gene. Insertion of putative or known activator binding sites upstream of the core promoter puts lacZ (beta-galactosidase) expression under the control of the corresponding activator. Although these constructs have proved to work well for most purposes, they have certain limitations: (1) they give significant and carbon-source-dependent lacZ background expression; (2) unlike most other yeast promoters, the CYC1 upstream region has a partially open chromatin structure with an accessible TATA box; (3) they use only a single, moderately sensitive reporter; and (4) the use of multicopy vectors can result in activator titration. Here, we introduce novel reporter plasmids based on the yeast MEL1 (alpha-galactosidase) gene that can overcome all of these limitations. It is also shown that background expression is due to fortuitous activator binding sites within the plasmid backbones that are insufficiently shielded from the core promoters in the common CYC1 reporter plasmids.

Binding Sites↗

Neighborhood-based tobacco advertising targeting adolescents.

Adolescent tobacco use remains a serious problem, and adolescents may be particularly receptive to the glamorous images tobacco companies use in advertisements. A relatively new form of neighborhood-based outdoor advertising, the illuminated bus-stop-shelter billboard, was studied to determine tobacco companies' use of this medium. We hypothesized that in 2 distinct San Francisco, California, neighborhoods, 1 predominantly white and the other mostly Latino, we would find a predominance of tobacco advertising on these billboards in both neighborhoods, that tobacco advertisements would be more prevalent in the minority Latino neighborhood, and that tobacco advertising would target adolescents in both neighborhoods. Each bus-stop-shelter billboard advertisement in the study areas from April 1992 to March 1993 was recorded. The type and frequency of products advertised and qualitative content of tobacco advertisements were analyzed. Adolescents' possible exposure to these advertisements was noted. Our main outcome measures were the percentage of tobacco advertising, possible adolescent exposure to this advertising, and themes of the tobacco advertisements. About 10% of all bus-stop-shelter billboard advertisements in each area promoted tobacco use. Possible exposures to these advertisements were greater in the Latino neighborhood because of a greater adolescent population. Qualitative analyses of tobacco advertisements suggested that adolescents are the primary targets. We urge physicians and educators to explicitly address this form of tobacco advertising, and we urge a ban on neighborhood-based tobacco advertising.

Adolescent↗