tailieunhanh - Báo cáo y học: " Rapid isolation of mycoviral double-stranded RNA from Botrytis cinerea and Saccharomyces cerevisiae"

Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành y học dành cho các bạn tham khảo đề tài: Rapid isolation of mycoviral double-stranded RNA from Botrytis cinerea and Saccharomyces cerevisiae | Castillo et al. Virology Journal 2011 8 38 http content 8 1 38 J VIROLOGY JOURNAL METHODOLOGY Open Access Rapid isolation of mycoviral double-stranded RNA from Botrytis cinerea and Saccharomyces cerevisiae Antonio Castillo Luis Cottet Miguel Castro Felipe Sepulveda Abstract Background In most of the infected fungi the mycoviruses are latent or cryptic the infected fungus does not show disease symptoms and it is phenotypically identical to a non-infected strain of the same species. Because of these properties the initial stage in the search for fungi infected with mycoviruses is the detection of their viral genome which in most of the described cases corresponds to double-stranded RNA dsRNA . So to analyze a large number of fungal isolates it is necessary to have a simple and rapid method to detect dsRNA. Results A rapid method to isolate dsRNA from a virus-infected filamentous fungus Botrytis cinerea and from a killer strain of Saccharomyces cerevisiae using commercial minicolumns packed with CF11 cellulose was developed. In addition to being a rapid method it allows to use small quantities of yeasts or mycelium as starting material being obtained sufficient dsRNA quantity that can later be analyzed by agarose gel electrophoresis treated with enzymes for its partial characterization amplified by RT-PCR and cloned in appropriate vectors for further sequencing. Conclusions The method yields high quality dsRNA free from DNA and ssRNA. The use of nucleases to degrade the DNA or the ssRNA is not required and it can be used to isolate dsRNA from any type of fungi or any biological sample that contains dsRNA. Background Mycoviruses or fungal viruses have properties that differentiate them from viruses that infect animals plants and bacteria 1-4 they do not infect intact cells and are transmitted vertically by intracellular routes meiosis and mitosis and horizontally by anastomosis of compatible hyphae or through sexual mating of yeast cells. Mycov-iruses