کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
8470747 | 1550012 | 2015 | 9 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Red fluorescent proteins for imaging Zymoseptoria tritici during invasion of wheat
دانلود مقاله + سفارش ترجمه
دانلود مقاله ISI انگلیسی
رایگان برای ایرانیان
کلمات کلیدی
Fpssdi1Tub2Mycosphaerella graminicolaRFPtdTomatomCherrymRFPeGFPSeptoria tritici blotchROIDPISample size - اندازهی نمونهdays post infection - روز پس از عفونتsuccinate dehydrogenase 1 - سوکین دد دهیدروژناز 1Protein localization - محلی سازی پروتئینregion of interest - منطقه مورد نظرenhanced green fluorescent protein - پروتئین فلورسنت سبز افزایش یافته استred fluorescent protein - پروتئین فلورسنت قرمزmonomeric red fluorescent protein - پروتئین فلورسنت قرمز مونومرFluorescent proteins - پروتئین های فلورسنتColocalization - کلوکالیزاسیونmonomeric cherry - گیلاس مونومر
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
بیولوژی سلول
پیش نمایش صفحه اول مقاله
چکیده انگلیسی
The use of fluorescent proteins (FPs) in plant pathogenic fungi provides valuable insight into their intracellular dynamics, cell organization and invasion mechanisms. Compared with green-fluorescent proteins, their red-fluorescent “cousins” show generally lower fluorescent signal intensity and increased photo-bleaching. However, the combined usage of red and green fluorescent proteins allows powerful insight in co-localization studies. Efficient signal detection requires a bright red-fluorescent protein (RFP), combined with a suitable corresponding filter set. We provide a set of four vectors, suitable for yeast recombination-based cloning that carries mRFP, TagRFP, mCherry and tdTomato. These vectors confer carboxin resistance after targeted single-copy integration into the sdi1 locus of Zymoseptoria tritici. Expression of the RFPs does not affect virulence of this wheat pathogen. We tested all four RFPs in combination with four epi-fluorescence filter sets and in confocal laser scanning microscopy, both in and ex planta. Our data reveal that mCherry is the RFP of choice for investigation in Z. tritici, showing highest signal intensity in epi-fluorescence, when used with a Cy3 filter set, and laser scanning confocal microscopy. However, mCherry bleached significantly faster than mRFP, which favors this red tag in long-term observation experiments. Finally, we used dual-color imaging of eGFP and mCherry expressing wild-type strains in planta and show that pycnidia are formed by single strains. This demonstrates the strength of this method in tracking the course of Z. tritici infection in wheat.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Fungal Genetics and Biology - Volume 79, June 2015, Pages 132-140
Journal: Fungal Genetics and Biology - Volume 79, June 2015, Pages 132-140
نویسندگان
M. Schuster, S. Kilaru, M. Guo, M. Sommerauer, C. Lin, G. Steinberg,