Vishal Gaurav - Nuclear DNA and protein content evaluation in Taxus plant cell cultures using multiparameter flow cytometry

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      Publication Details (including relevant citation   information):

      Authors: Vishal Gaurav, Amit Shavit, Susan C. Roberts

      Publication date: 2010

      Conference name: The American Chemical Society (ACS) Proceedings

      Publisher: The ACS National Meeting, San Francisco, CA

      Abstract:

      Plant cell cultures of Taxus provide the most reliable production   methods for the anti-cancer drug paclitaxel. In order to   comprehend the inherent culture heterogeneity and production   variability in cell cultures, it is essential that the cellular   metabolism is studied at the genomic level. Genomic stability in   plant cell cultures is crucial as it affects cell growth and   division, metabolite accumulation and protein synthesis. A rapid   and efficient method to prepare nuclei suspensions from   aggregated cell cultures of Taxus was employed. Methods were   subsequently developed to simultaneously stain them for DNA and   protein content using Propidium Iodide and Fluorescein   Isothiocyanate respectively. Flow cytometry was used to analyze   and quantify the DNA content and genome size of Taxus using known   reference species as standards. Furthermore, their genomic   stability was evaluated by correlating DNA content and genome   size with cell size and complexity, protein content, and   elicitation effects using multiparameter flow cytometry. These   techniques to evaluate and correlate various culture   characteristics can be very useful in designing superior bio   processes for enhanced production.

      Address (URL): https://sites.google.com/site/vishalpubs/Nuclear_DNA_and_protein_content_evaluat ion_in_Taxus_plant_cell_cultures_using_multiparameter_flow_cytometry.pdf