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METHOD AND KIT FOR CRYOPRESERVING XENOPUS LAEVIS EGG EXTRACT, CONCENTRATED XENOPUS LAEVIS EGG EXTRACT, AND METHOD FOR ANALYZING CELL CYCLE

Foreign code F180009513
File No. (S2017-0321-N0)
Posted date Nov 2, 2018
Country WIPO
International application number 2018JP002546
International publication number WO 2018139601
Date of international filing Jan 26, 2018
Date of international publication Aug 2, 2018
Priority data
  • P2017-014441 (Jan 30, 2017) JP
Title METHOD AND KIT FOR CRYOPRESERVING XENOPUS LAEVIS EGG EXTRACT, CONCENTRATED XENOPUS LAEVIS EGG EXTRACT, AND METHOD FOR ANALYZING CELL CYCLE
Abstract A method for cryopreserving a Xenopus laevis egg extract, characterized by comprising a partition step for partitioning the Xenopus laevis egg extract into water and solid matters and a freezing step for separately freezing the water and the solid matters.
Outline of related art and contending technology BACKGROUND ART
Xenopus egg extracts is, the highly tuned to the cell-free cell cycle (cell-free extracts) the only experimental system can be realized.In this experimental system, using an antibody specific for a particular factor may be removed from the system and cell cycle progression (immune removed) by the control, as well as DNA replication and distribution control of the factors necessary for the unambiguous identification.
In addition, in cell cycle control and understanding of the mechanism, by using a yeast single powerful means and research, genes involved in these processes have been identified in turn.However, the eggs Xehopiis using extract cell-free system, as compared to yeast excellent in the following points. (1) Biochemical, physiological studies are suitable: for example, entity M phase promoting factor of the complex MPF cyclinB/Cdk1, Xenopus egg extracts used by the cell-free system, have been identified for the first time (for example, see Non-Patent Document 1.). (2) In vitro in the partial reconstruction. (3) Different yeast and multicellular animal and the frog is, closer to the human model.To this, the cell-free system using Xenopus egg extracts by, the cell cycle specific multicellular animal control mechanism has been revealed.
Scope of claims (In Japanese)請求の範囲
[請求項1]
 アフリカツメガエル卵抽出液から水分と固形分とを分画する分画工程と、
 前記水分及び前記固形分を別々に冷凍する冷凍工程と、を備えることを特徴とするアフリカツメガエル卵抽出液の冷凍保存方法。
[請求項2]
 前記分画工程において、分画分子量が1000以上のろ過膜を用いる請求項1に記載のアフリカツメガエル卵抽出液の冷凍保存方法。
[請求項3]
 前記分画工程において、分画分子量が10000以上のろ過膜を用いる請求項1又は2に記載のアフリカツメガエル卵抽出液の冷凍保存方法。
[請求項4]
 前記冷凍工程において、-0.5℃/分以上-1.5℃/分以下の速度で前記水分及び前記固形分を冷凍する請求項1~3のいずれか一項に記載のアフリカツメガエル卵抽出液の冷凍保存方法。
[請求項5]
 水分分画用ろ過膜を備える遠心チューブを含むことを特徴とするアフリカツメガエル卵抽出液の冷凍保存用キット。
[請求項6]
 前記水分分画用ろ過膜の分画分子量が1000以上である請求項5に記載のアフリカツメガエル卵抽出液の冷凍保存用キット。
[請求項7]
 前記水分分画用ろ過膜の分画分子量が10000以上である請求項5又は6に記載のアフリカツメガエル卵抽出液の冷凍保存用キット。
[請求項8]
 タンパク質濃度が83mg/mL以上103mg/mL以下であり、
 分子量が20000以上30000以下のタンパク質の含有量が、総タンパク質量に対して、20質量%以下であり、且つ、
 分子量が100000以上150000以下のタンパク質の含有量が、総タンパク質量に対して、3質量%以上であることを特徴とするアフリカツメガエル卵濃縮抽出液。
[請求項9]
 請求項1~4のいずれか一項に記載のアフリカツメガエル卵抽出液の冷凍保存方法の工程で冷凍保存されたアフリカツメガエル卵抽出液の水分及び固形分を解凍し、混合して再構築されたアフリカツメガエル卵抽出液を用いることを特徴とする細胞周期の分析方法。
  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • INTER-UNIVERSITY RESEARCH INSTITUTE CORPORATION RESEARCH ORGANIZATION OF INFORMATION AND SYSTEMS
  • Inventor
  • SHIMAMOTO Yuta
  • TAKAGI Jun
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JO JP KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG

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