Localization of DNA and protein in Tipula iridescent virus (TIV) by enzymatic digestion and electron microscopy.

R S THOMAS, R C WILLIAMS
{"title":"Localization of DNA and protein in Tipula iridescent virus (TIV) by enzymatic digestion and electron microscopy.","authors":"R S THOMAS,&nbsp;R C WILLIAMS","doi":"10.1083/jcb.11.1.15","DOIUrl":null,"url":null,"abstract":"<p><p>De-embedded ultrathin sections of ethanol-fixed Tipula Iridescent Virus particles were incubated with pepsin at pH 1.8, trypsin at pH 7.7, and DNase at pH 7.7. The outer shell of the particles, but not an inner core, was removed by the action of pepsin. Conversely, the inner core, but not the outer shell, was removed by the action of trypsin and DNase in combination, but not by either enzyme acting alone. These results are taken to mean that the outer shell of the particles is protein in nature and the inner core is nucleoprotein. Whole virus particles were also exposed to the same 3 enzymes. Trypsin and/or DNase had no effect on the whole particles, while pepsin at pH 1.8 digested away the outer shell of the particles and released an intact core, resistant to pepsin. The protein nature of the digested outer shells and the nucleoprotein nature of the released cores were confirmed by ultraviolet absorption spectra. Chemical analyses showed that the cores contain 89 per cent of the whole virus phosphorus but only 35 per cent of the nitrogen, while the outer shells contain only 5 per cent of the phosphorus but 63 per cent of the nitrogen. On the basis of nitrogen: phosphorus ratios the composition of the cores is estimated to be about 30 per cent DNA and 60 to 65 per cent protein.</p>","PeriodicalId":22618,"journal":{"name":"The Journal of Biophysical and Biochemical Cytology","volume":"11 ","pages":"15-29"},"PeriodicalIF":0.0000,"publicationDate":"1961-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1083/jcb.11.1.15","citationCount":"19","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"The Journal of Biophysical and Biochemical Cytology","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1083/jcb.11.1.15","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 19

Abstract

De-embedded ultrathin sections of ethanol-fixed Tipula Iridescent Virus particles were incubated with pepsin at pH 1.8, trypsin at pH 7.7, and DNase at pH 7.7. The outer shell of the particles, but not an inner core, was removed by the action of pepsin. Conversely, the inner core, but not the outer shell, was removed by the action of trypsin and DNase in combination, but not by either enzyme acting alone. These results are taken to mean that the outer shell of the particles is protein in nature and the inner core is nucleoprotein. Whole virus particles were also exposed to the same 3 enzymes. Trypsin and/or DNase had no effect on the whole particles, while pepsin at pH 1.8 digested away the outer shell of the particles and released an intact core, resistant to pepsin. The protein nature of the digested outer shells and the nucleoprotein nature of the released cores were confirmed by ultraviolet absorption spectra. Chemical analyses showed that the cores contain 89 per cent of the whole virus phosphorus but only 35 per cent of the nitrogen, while the outer shells contain only 5 per cent of the phosphorus but 63 per cent of the nitrogen. On the basis of nitrogen: phosphorus ratios the composition of the cores is estimated to be about 30 per cent DNA and 60 to 65 per cent protein.

用酶解和电镜技术定位虹彩蒂普拉病毒(TIV)的DNA和蛋白质。
将脱包体超薄切片用乙醇固定Tipula彩虹病毒颗粒,与pH 1.8的胃蛋白酶、pH 7.7的胰蛋白酶和pH 7.7的dna酶孵育。颗粒的外壳,而不是内核,被胃蛋白酶的作用去除。相反,内核,而不是外壳,被胰蛋白酶和dna酶联合作用,而不是单独作用。这些结果被认为意味着粒子的外壳本质上是蛋白质,内核是核蛋白。整个病毒颗粒也暴露在同样的3种酶中。胰蛋白酶和/或DNase对整个颗粒没有影响,而pH为1.8的胃蛋白酶消化掉了颗粒的外壳,释放出一个完整的核,对胃蛋白酶具有抗性。用紫外吸收光谱测定了酶解壳的蛋白质性质和释放核的核蛋白性质。化学分析表明,病毒核含有整个病毒磷的89%,但只含有35%的氮,而外壳只含有5%的磷,但含有63%的氮。根据氮磷比,岩心的组成估计约为30%的DNA和60%至65%的蛋白质。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信