caa a22 4500 397494572 CHVBK 20180308164518.0 cr unu---uuuuu 161202e199503 xx s 000 0 eng 10.1093/oxfordjournals.jbchem.a124732 doi (NATIONALLICENCE)oxford-10.1093/oxfordjournals.jbchem.a124732 An Improved Method for Extraction of mRNA and Protein from the Fungus Chaetomium gracile with Anhydrous Hydrogen Fluoride [Elektronische Daten] [Satoru Oshiro, Nobuhiko Katsura] We previously reported that DNAs directly applicable to restriction analyses and transformation of Escherichia coli can be extracted from fungi and yeasts by use of anhydrous hydrogen fluoride (HF) under a mild condition, 5 min at 0°C [Oshiro, S., Katsura, N., Kitada, K., and Gunge, N. (1987) FEBS Lett. 220, 383-386]. In the present investigation, we examined whether this improved method is also applicable to extraction of RNA and protein from the fungus Chaetomium gracile. The RNA and protein were effectively extracted from the fungus after anhydrous hydrogen fluoride (HF) treatment for a short time (1 min) at 0°C. The extracted poly (A)-enriched mRNA and proteins were fully intact: the mRNA purified by messenger-activated paper with poly(U) directed not only the incorporation of [3H]glycine into polypeptides but also the synthesis in a rabbit reticulo-cyte lysate cell-free system of proteins reactive to antibodies against the soluble fraction extracted from the fungus by HF. Analyses by gel filtration and polyacrylamide electro-phoresis under native conditions showed that dextranase extracted from the fungus by HF under the same conditions had the same molecular weight and electrophretic mobility as the enzyme excreted into the medium. This suggests that the mRNAs and proteins extracted by this method are also applicable to protein synthesis directed in a cell-free system and to enzyme purification from a fungus insusceptible to lytic enzymes. This method provides a pure preparation of mRNA within 5 h and starting materials for protein purification wthin lh. © 1995 BY THE JOURNAL OF BIOCHEMISTRY Regular Papers nationallicence anhydrous hydrogen fluoride nationallicence protein extraction nationallicence RNA extraction nationallicence Oshiro Satoru Department of Biochemical Genetics, Medical Research Institute, Tokyo Medical and Dental University 1-5-45 Yushima, Bunkyo-ku, Tokyo 113 aut Katsura Nobuhiko Department of Oral Biochemistry, School of Dentistry, Nagasaki University, 7-1 Sakamoto-machi, Nagasaki 852 aut The Journal of Biochemistry Oxford University Press 117/3(1995-03), 475-479 0021-924X 117:3<475 1995 117 jbchem https://doi.org/10.1093/oxfordjournals.jbchem.a124732 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1093/oxfordjournals.jbchem.a124732 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Oshiro Satoru Department of Biochemical Genetics, Medical Research Institute, Tokyo Medical and Dental University 1-5-45 Yushima, Bunkyo-ku, Tokyo 113 aut NATIONALLICENCE 700 1- Katsura Nobuhiko Department of Oral Biochemistry, School of Dentistry, Nagasaki University, 7-1 Sakamoto-machi, Nagasaki 852 aut NATIONALLICENCE 773 0- The Journal of Biochemistry Oxford University Press 117/3(1995-03), 475-479 0021-924X 117:3<475 1995 117 jbchem Metadata rights reserved CC BY-NC-4.0 http://creativecommons.org/licenses/by-nc/4.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-oxford