INHIBITION OF PROTEIN SYNTHESIS IN RABBIT RETICULOCYTE BY NIVALENOL A TOXIC PRINCIPLE ISOLATED FROM FUSARIUM NIVALE-GROWING RICE
INHIBITION OF PROTEIN SYNTHESIS IN RABBIT RETICULOCYTE BY NIVALENOL A TOXIC PRINCIPLE ISOLATED FROM FUSARIUM NIVALE-GROWING RICE
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DOI：
10.1093/oxfordjournals.jbchem.a128919
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发表时间：
1968-01-01
影响因子：
2.7
通讯作者：
TATSUNO, T
中科院分区：
文献类型：
--
作者：
UENO, Y;HOSOYA, M;TATSUNO, T
方法网织红细胞兔于第1~4天皮下注射1.2%苯肼溶液5 mg/kg，第7天心脏穿刺法放血。血液在2000g中保存5min，网织红细胞含量在85%以上的小球用冰盐水洗两次，然后悬浮在两体积的生理盐水中。网织红细胞在本报告中被称为“全细胞”，用于毒性原理的生物测定，如下所述。细胞在冷水浴中保存，两天后仍有活力。全细胞生物测定--在总体积为0.9ml的洛克-林格液0.6ml、全细胞0.2ml、试验液0.1ml中，将溶液在37·Ž的摇床中预培养15min，加入0.1ml(0.1mC)的1-14C-亮氨酸(13mC/mm)继续孵育40min。反应结束后，加入1.0ml 10%高氯酸停止反应，沉淀用冰冷的5%高氯酸洗涤三次，用乙醇：乙醚(体积比3：1)洗涤两次。用气体流量计数器计算洗涤后沉淀物的放射性活度。
METHODS Reticulocyte-Rabbits were injected subcutaneously with 5mg/kg of 1.2% phenylhydrazine dissolved in 0.9% normal saline on days 1-4, and on day 7 they were bled by cardiac puncture. The blood was cen trifuged at 2000g for 5 min and the pellet, consisting of more than 85% reticulocytes, was washed twice with an ice-cold saline and then suspend in two volumes of the saline. The reticulocyte was referred as" whole cell" in this report and employed for the bioassay of the toxic principle as described below. When the cells were stored in an ice-cold bath, they were active even after two days.Bioassay of the Toxic Principle with Whole Cells-For the bioassay of the toxin in 0.6 ml of Locke-Ringer's solution, 0.2 ml whole cell, 0.1 ml test solution, in a total volume of 0.9 ml, the solution was preincubated at 37• Ž for 15 min in a shaking incubater, and 0.1 ml (0.1 ƒÊC) of 1-14C-leucine(13mC/mM) was added to continue the incubation for 40 min. After the incu bation, the reaction was stopped by the addition of 1.0 ml of 10% perchloric acid, and the precipitate was washed three times with ice-cold 5% perchloric acid, twice with ethanol: ether (3: 1, by vol.). The radio activity of the washed precipitate was counted by a gas flow counter.