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Park, Gyungsoon,Borkovich, Katherine A Humana Press 2012 METHODS IN MOLECULAR BIOLOGY -CLIFTON THEN TOTOWA- Vol.883 No.-
<P>Due to crucial roles in gene regulation, noncoding small RNAs (smRNAs) of 20-30 nucleotides (nt) have been intensively studied in mammals and plants, and are known to be implicated in significant diseases and metabolic disorders. Elucidation of biogenesis mechanisms and functional characterization of smRNAs are often achieved using tools, such as separation of small-sized RNA and high-throughput sequencing. Although RNA interference pathways such as quelling and meiotic silencing have been well described in Neurospora crassa, knowledge of smRNAs in filamentous fungi is still limited compared to other eukaryotes. As a prerequisite for study, isolation and sequence analysis of smRNAs are necessary. We developed a protocol for isolation and library construction of smRNAs of 20-30 nt for Solexa sequencing in two -filamentous fungi, N. crassa and Fusarium oxysporum f.sp. lycopersici. Using 200-300 관g total RNA, smRNA was isolated by size fractionation, ligated with adapters, and amplified by RT-PCR for Solexa sequencing. Sequence analysis of several cDNA clones showed that the cloned smRNAs were not tRNAs and rRNAs and were fungal genome specific.</P>
Lee, Geon Joon,Park, Gyungsoon,Choi, Eun Ha 한국물리학회 2017 THE JOURNAL OF THE KOREAN PHYSICAL SOCIETY Vol. No.
<P>We studied the effect of plasma treatment on the optical, structural and biological properties of Neurospora crassa (N. crassa) spores. An atmospheric-pressure plasma jet (APPJ) was used to generate reactive oxygen and nitrogen species in aqueous solution. The APPJ treatment of N. crassa spores in water significantly reduced the viability of spores. The reduction in the spore viability can be attributed to the reactive species from the plasma itself and those derived from the reaction of plasma radicals with aqueous solution. These structural modifications were contingent on the medium in which N. crassa spores were suspended; plasma treatment of N. crassa spores in PBS did not significantly affect the viability of spores as compared with N. crassa spores in water. Scanning electron microscopy images and circular dichroism spectra indicated that the spore cell wall was damaged by plasma treatment. The optical absorption spectrum of untreated N. crassa spores exhibited two resonance absorption bands at approximately lambda(1) ae 260 nm and lambda(2) ae 472 nm, originating from deoxyribonucleic acid (DNA) and beta-carotene. The Raman spectrum of untreated N. crassa spores exhibited three main peaks at 1519, 1157 and 1006 cm (-1), attributed to beta-carotene inside the cell wall. The Raman spectra showed that the APPJ treatment of N. crassa spores in water caused degradation of beta-carotene, affecting the viability of spores.</P>