کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
8085677 1521800 2018 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Improved DNA/protein delivery in microalgae - A simple and reliable method for the prediction of optimal electroporation settings
موضوعات مرتبط
مهندسی و علوم پایه مهندسی انرژی انرژی های تجدید پذیر، توسعه پایدار و محیط زیست
پیش نمایش صفحه اول مقاله
Improved DNA/protein delivery in microalgae - A simple and reliable method for the prediction of optimal electroporation settings
چکیده انگلیسی
Genetic transformation of microalgae remains a challenge due to poor intracellular delivery of exogenous molecules. This limitation is caused by the structure and composition of the cell wall and cell membrane of each species. Moreover, successful delivery of proteins or nucleic acids cannot be assessed by determining transformability since their functionality is not always known in the studied microorganism. We propose a quick and effective screening tool for the prediction and optimization of electroporation settings by monitoring cell permeability and viability using Sytox Green and propidium iodide respectively. We determined voltage settings for the microalgae Chlamydomonas reinhardtii, Chlorella vulgaris, Neochloris oleoabundans and Acutodesmus obliquus. To evaluate the predicted settings, we delivered labelled DNA and proteins into the cells. We demonstrated that high transformation efficiencies can be accomplished when predicted values were applied with functional plasmids. Additionally, we increased transformation efficiencies by testing cell concentrations, light intensities and fragment sizes. This method can be used to determine suitable transformation conditions for non-transformed microalgae species and to increase the insight on established transformation protocols.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Algal Research - Volume 33, July 2018, Pages 448-455
نویسندگان
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