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＜departmental bulletin paper＞
On an Isolation Method of Protein from Single Cell by Impact-Cell-Mill (ICM) (Part. 4 ) : Isolation of Nucleic Acid and Protein from Single Cell

Creator
Creator Name
早川, 功
Hayakawa, Isao
Affiliation
Affiliation Name
九州大学農学部食品製造工学教室
Laboratory of Food Technology, Faculty of Agriculture, Kyushu University
Creator Name
安松, 睦郎
Yasumatsu, Mutsuo
Affiliation
Affiliation Name
九州大学農学部食品製造工学教室
Laboratory of Food Technology, Faculty of Agriculture, Kyushu University
Creator Name
野村, 男次
Nomura, Danji
Affiliation
Affiliation Name
九州大学農学部食品製造工学教室
Laboratory of Food Technology, Faculty of Agriculture, Kyushu University
Language
Japanese
Publisher
九州大學農學部
Faculty of Agriculture, Kyushu University
Date
1973-07
Source Title
Science bulletin of the Faculty of Agriculture, Kyushu University
Vol
27
Issue
3/4
First Page
139
Last Page
146
Publication Type
Version of Record
Access Rights
open access
JaLC DOI
https://doi.org/10.15017/23135
Related DOI
九州大學農學部學藝雜誌 || 27(3/4) || p139-146
Science bulletin of the Faculty of Agriculture, Kyushu University || 27(3/4) || p139-146
http://www.agr.kyushu-u.ac.jp/
Related URI
九州大學農學部學藝雜誌 || 27(3/4) || p139-146
Science bulletin of the Faculty of Agriculture, Kyushu University || 27(3/4) || p139-146
http://www.agr.kyushu-u.ac.jp/
Relation
九州大學農學部學藝雜誌 || 27(3/4) || p139-146
Science bulletin of the Faculty of Agriculture, Kyushu University || 27(3/4) || p139-146
http://www.agr.kyushu-u.ac.jp/

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Details

Record ID
23135
Peer-Reviewed
Unrefereed
ISSN
0368-6264
NCID
AN0005519X
Notes
An experiment was carried out to establish a new method for the isolation of protein and nucleic acid from single cell employing an Impact-Cell-Mill. Chemical pretreatment with 0.7 N perchloric acid gave a good result to isolate nucleic acid from Baker's yeast. About 85 % of protein was obtained using an Impact-Cell-Mill after the treatment of 0.7 N perchloric acid. Concurrent treatment of an Impact-Cell-Mill and chemical procedure gave also a good result. In this case, 0.7 N perchloric acid and 40 γ/ml of ribonuclease were used, and about 70% of nucleic acid was isolated and 85 % of protein was recovered with the treatment of an Impact-Cell-Mill. The third is an utilization of an Impact-Cell-Mill as a reactor. In this case, adding 180 γ/ml of ribounclease, nucleic acid was isolated about 90 % and 95 % of protein was obtained from the supernatant of yeast suspension treated with an Impact-Cell-Mtll.
酵母など単細胞生物からタンパク質ならびに核酸類を連続的かつ大量に分離する方法の開発を目的として,Impact－Cell－Mill（ICM）と化学的な処理方法を組合せ（併用し）て,次のような結果を得た.1）化学的な前処理とICM処理の組合せにより,核酸類とタンパク質の分離回収する方法として過塩素酸処理が良好であった.0.7N過塩素酸処理で核酸類の約85％が抽出可能となり,抽出残渣に0.5％NaOHを添加して,ICM処理を施した結果,タンパク態窒素の約85％が回収出来た.2）化学的作用とICM処理の併用の場合.生酵母懸濁液に過塩素酸または核酸分解酵素を添加し,ICM処理を施すと,核酸類の約70％が5％TCA可溶性となり,37℃,20分間振盤抽出した場合に比較して約70倍以上の値を示した.また,タンパク態窒素量の約85％が回収出来た.3）ICMと化学的後処理の場合.ICM処理で得られた生酵母破砕液の上澄液に過塩素酸または核酸分解酵素を添加した衝再びICM処理を完施した結果,2N過塩素酸で92.3％,リボヌクレアーゼで89.8％がTCA可溶性リンとなり上澄みに移行し,タンパク態窒素量の約95％が沈澱として回収された.
Created Date 2012.07.25
Modified Date 2022.11.11

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