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噬菌体T7和T7衣壳在琼脂糖凝胶中的电泳

Electrophoresis of bacteriophage T7 and T7 capsids in agarose gels.

作者信息

Serwer P, Pichler M E

出版信息

J Virol. 1978 Dec;28(3):917-28. doi: 10.1128/JVI.28.3.917-928.1978.

Abstract

Agarose gel electrophoresis of the following was performed in 0.05 M sodium phosphate-0.001 M MgCl2 (pH 7.4): (i) bacteriophage T7; (ii) a T7 precursor capsid (capsid I), isolated from T7-infected Escherichia coli, which has a thicker and less angular envelope than bacteriophage T7; (iii) a second capsid (capsid II), isolated from T7-infected E. coli, which has a bacteriophage-like envelope; and (iv) capsids (capsid IV) produced by temperature shock of bacteriophage T7. Bacteriophage T7 and all of the above capsids migrated towards the anode. In a 0.9% agarose gel, capsid I had an electrophoretic mobility of 9.1 +/- 0.4 X 10(-5) cm2/V.s; bacteriophage T7 migrated 0.31 +/- 0.02 times as fast as capsid I. The mobilities of different preparations of capsid II varied in such gels: the fastest-migrating capsid II preparation was 0.51 +/- 0.03 times as fast as capsid I and the slowest was 0.37 +/- 0.02 times as fast as capsid I. Capsid IV with and without the phage tail migrated 0.29 +/- 0.02 and 0.42 +/- 0.02 times as fast as capsid I. The results of the extrapolation of bacteriophage and capsid mobilities to 0% agarose concentration indicated that the above differences in mobility are caused by differences in average surface charge density. To increase the accuracy of mobility comparisons and to increase the number of samples that could be simultaneously analyzed, multisample horizontal slab gels were used. Treatment with the ionic detergent sodium dodecyl sulfate converted capsid I to a capsid that migated in the capsid II region during electrophoresis through agarose gels. In the electron microscope, most of the envelopes of these latter capsids resembled the capsid II envelope, but some envelope regions were thicker than the capsid II envelope.

摘要

在0.05M磷酸钠 - 0.001M氯化镁(pH 7.4)中对以下物质进行琼脂糖凝胶电泳:(i)噬菌体T7;(ii)从T7感染的大肠杆菌中分离出的T7前衣壳(衣壳I),其包膜比噬菌体T7更厚且棱角更少;(iii)从T7感染的大肠杆菌中分离出的第二种衣壳(衣壳II),其具有类似噬菌体的包膜;以及(iv)通过噬菌体T7温度休克产生的衣壳(衣壳IV)。噬菌体T7和上述所有衣壳都向阳极迁移。在0.9%琼脂糖凝胶中,衣壳I的电泳迁移率为9.1±0.4×10⁻⁵cm²/V·s;噬菌体T7的迁移速度是衣壳I的0.31±0.02倍。不同制备的衣壳II在这种凝胶中的迁移率有所不同:迁移最快的衣壳II制剂的速度是衣壳I的0.51±0.03倍,最慢的是衣壳I的0.37±0.02倍。带有和不带有噬菌体尾部的衣壳IV的迁移速度分别是衣壳I的0.29±0.02倍和0.42±0.02倍。将噬菌体和衣壳迁移率外推至0%琼脂糖浓度的结果表明,上述迁移率差异是由平均表面电荷密度差异引起的。为了提高迁移率比较的准确性并增加可同时分析的样品数量,使用了多样品水平平板凝胶。用离子去污剂十二烷基硫酸钠处理后,衣壳I在通过琼脂糖凝胶电泳时转变为在衣壳II区域迁移的衣壳。在电子显微镜下,这些后一种衣壳的大多数包膜类似于衣壳II包膜,但一些包膜区域比衣壳II包膜更厚。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/42c5/525816/f02d92958e9d/jvirol00204-0262-a.jpg

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