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(023) Atrazine-mediated alterations of gene expression in Chironomus tentans (Diptera: Chironomidae).
Anderson, Troy*,1, Jin-Clark, Ying1, Zhu, Kun Yan1, 1 Department of Entomology, Kansas State University, Manhattan, Kansas
ABSTRACT- A novel molecular technique called restriction fragment differential display-PCR (RFDD-PCR) was used to determine altered gene expression in fourth-instar larvae of the aquatic midge, Chironomus tentans (Diptera: Chironomidae), exposed to the triazine herbicide atrazine. Using specific primers complementary to adaptor sequences, cDNA restriction fragments from control and atrazine-treated C. tentans were amplified by PCR. Differentially expressed cDNA fragments were individually subcloned and sequenced. Sequence comparisons revealed a number of over- and under-expressed genes in atrazine-treated C. tentans. The identities of selected genes, with altered expression, were determined by cloning and sequencing their full-length cDNAs. The alteration of gene expression was further verified by Northern blot analysis. The results of this study suggest that RFDD-PCR is a useful approach to study atrazine-mediated alterations of gene expression in C. tentans.
Key words: atrazine, gene expression, RFDD-PCR, Chironomus tentans
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