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分离的胃黏膜细胞合成的放射性标记蛋白质的计算分辨率和相对比放射性。

Computed resolution and relative specific radioactivities of radiolabelled proteins synthesized by isolated gastric mucosal cells.

作者信息

Trotman C N, Greenwell J R

出版信息

Biochem J. 1979 Jan 15;178(1):159-64. doi: 10.1042/bj1780159.

Abstract
  1. By radiolabelling, polyacrylamide-gel electrophoresis in the presence of sodium dodecyl sulphate and fluorography, more than 50 soluble proteins in the molecular-weight range 15000-100000 were shown to continue to be synthesized after cells had been isolated from rat gastric mucosa. 2. Densitometric measurements of stained gels and fluorographic films were processed by computer to resolve individual overlapping Gaussian peaks corresponding to the protein bands. 3. Comparison of resolved peak areas of radioactivity and staining showed certain bands to have characteristically high relative specific radioactivities. 4. The computer programs (in FORTRAN) permit the analysis of a single densitometric trace or the simultaneous comparison of a corresponding pair of densitometric records of stained gels, or of fluorographic films, or a combination. Central processing unit time is used economically. 5. The programs identify the Gaussian components that contribute to the records and estimate their means, standard deviations and enclosed areas. These estimates are improved by a piecewise iterative method that minimizes the errors between the calculated and the experimental data. 6. Relative specific radioactivities are calculated as the normalized ratio of the area of a fluorographic film peak and the area of the corresponding stained gel peak. The computer programs have been deposited as Supplementary Publication SUP 50094 (55 pages) at the British Library Lending Division, Boston Spa, Wetherby, West Yorkshire LS23 7BQ, U.K., from whom copies can be obtained on the terms given in Biochem. J. (1978) 169, 5.
摘要
  1. 通过放射性标记、在十二烷基硫酸钠存在下的聚丙烯酰胺凝胶电泳以及荧光自显影,结果显示,从大鼠胃黏膜分离出细胞后,分子量在15000至100000范围内的50多种可溶性蛋白质仍在继续合成。2. 对染色凝胶和荧光自显影片进行光密度测量,并通过计算机处理,以分辨与蛋白条带相对应的各个重叠的高斯峰。3. 对放射性和染色的分辨峰面积进行比较,结果显示某些条带具有特征性的高相对比放射性。4. 计算机程序(用FORTRAN语言编写)可对单个光密度曲线进行分析,也可同时比较染色凝胶、荧光自显影片或两者组合的相应一对光密度记录。中央处理器时间得到了有效利用。5. 这些程序可识别对记录有贡献的高斯成分,并估计它们的均值、标准差和封闭面积。通过一种分段迭代方法可改进这些估计值,该方法可使计算数据与实验数据之间的误差最小化。6. 相对比放射性计算为荧光自显影片峰面积与相应染色凝胶峰面积的归一化比值。这些计算机程序已作为补充出版物SUP 50094(共55页)存放在英国西约克郡韦瑟比波士顿温泉市大英图书馆出借部,其存档编号为LS23 7BQ,可按《生物化学杂志》(1978年)第169卷第5期所述条件从该处获取副本。
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9464/1186492/0da038f16135/biochemj00468-0164-a.jpg

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本文引用的文献

1
RAT STOMACH PREPARATION IN VITRO.
Biochem Pharmacol. 1965 Mar;14:355-7. doi: 10.1016/0006-2952(65)90202-9.
10
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Cell Biol Int Rep. 1977 Nov;1(6):541-7. doi: 10.1016/0309-1651(77)90092-3.

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