An immunoassay has been developed for trichlorophenol analysis on the basis of theoretical chemistry modeling studies. These data have allowed us to choose the optimum chemical structure of the immunizing hapten according to realistic similarities with the target analyte. The synthesis of this hapten and the subsequent application of an appropriate immunization protocol have lead to the production of polyclonal antibodies against the target analyte. A homologous direct competitive ELISA has been developed that can be carried out in about 1 h. It has a limit of detection of 0.2 +/- 0.06 microg/L (1.01 +/- 0.3 nM) and it has been proven to tolerate a wide range of ionic strengths and pH values. Thus, the assay has acceptable features in samples with ionic strength between 4 and 56 mS/cm and pH values between 5.5 and 9.5. Studies on the selectivity of this immunoassay have demonstrated a high recognition of the corresponding brominated analogues. Other phenolic compounds do not interfere significantly in the analysis of 2,4,6-trichorophenol using this immunochemical technique. The accuracy of the assay has been evaluated using certified and spiked samples.
The sex pheromone of the female processionary moth, Thaumetopoea pityocampa, is a unique C16 enyne acetate that is biosynthesized from palmitic acid. Three consecutive desaturation reactions transform this saturated precursor into the triunsaturated fatty acyl intermediate: formation of (Z)-11-hexadecenoic acid, acetylenation to 11-hexadecynoic acid, and final ⌬ 13 desaturation to (Z)-13-hexadecen-11-ynoic acid. By using degenerate primers common to all reported insect desaturases, a single cDNA sequence was isolated from total RNA of T. pityocampa female pheromone glands. The full-length transcript of this putative desaturase was expressed in elo1⌬/ole1⌬ yeast mutants (both elongase 1 and ⌬ 9 desaturase-deficient) for functional assays. The construct fully rescued the ⌬ole1 yeast phenotype, confirming its desaturase activity. Analysis of the unsaturated products from transformed yeast extracts demonstrated that the cloned enzyme showed ⌬ 11 desaturase, ⌬ 11 acetylenase, and ⌬ 13 desaturase activities. Therefore, this single desaturase may account for the three desaturation steps involved in the sex pheromone biosynthetic pathway of the processionary moth.acetylene ͉ cloning ͉ enyne ͉ Thaumetopoea pityocampa ͉ yeast
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