CONSTRUCTION OF MOLECULAR CATCHERS FOR SHORT NON-CODING RNA (SHORT NCRNA) USING MUTANT RIBONUCLEASE 1-AND PAZ DOMAIN-DISPLAYING YEASTS
Accelerating discovery of novel functions of non-coding RNAs (ncRNAs) suggests that one-time junk sequences found in genome have critical roles in cellular process. Now there is a need to clarify the entire component ncRNAs as well as other cellular components which exist in specific biological phase, and to indicate how they play a role in biological system. Here we constructed protein-based molecular catcher for short ncRNAs, and tried to separate short RNAs using HPLC. For ncRNA catcher, we used mutant human ribonuclease 1 (hRNasel) protein which has no RNA cleavage activities, and PAZ domain derived from human argonaute 2 (hAgo2) protein which has micro RNA (miRNA)-binding ability. Using these molecular catchers, we estimated short RNA-binding and -releasing abilities and calculated the efficiency. Furthermore, we estimated the separation ability and detectability of short RNAs using HPLC. Our method will have, when coupled with sequence determination using MS or MS/ MS, a potential to reveal all the component ncRNAs that play a role in complex web of interactions in cellular systems.
Short non-coding RNA (short ncRNA) yeast molecular catcher surface display system mutant RNasel PAZ domain
Natsuko Miura Eri Hayashi Hironobu Morisaka Kouichi Kuroda Mitsuyoshi Ueda
Division of Applied Life Sciences, Graduate School of Agriculture, Kyoto University Sakyo-ku, Kyoto 606-8502, Japan
国际会议
The 6th International Forum on Post-genome Technologies(6IFPT)(第六届国际后基因组生命科学技术学术论坛)
北京
英文
245-248
2009-09-17（万方平台首次上网日期，不代表论文的发表时间）