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用二乙氨基乙基葡聚糖增强昆虫细胞单层中的杆状病毒蚀斑试验

Enhancement of baculovirus plaque assay in insect cell monolayers by DEAE-dextran.

作者信息

Carrascosa A L

机构信息

Centro de Biología Molecular (C.S.I.C.-U.A.M.), Facultad de Ciencias, Universidad Autónoma de Madrid, Spain.

出版信息

Biotechniques. 1994 Jun;16(6):1078-81, 1083-5.

PMID:8074873
Abstract

The presence of DEAE-dextran in the agarose overlay, when titrating wild-type or recombinant baculoviruses by plaque assay, resulted in a higher definition and contrast of viral plaques and increased the plaque number and mean diameter at least by a factor of 2x on day 5 post infection. This increase was related to neither a larger production of infectious virus nor of recombinant protein and did not occur when the polycation was only present in the virus inoculum or when insect cell monolayers were preincubated for 60 min with it. The extension of the observations to a number of different recombinant viruses from varied sources, including several baculoviruses that could not consistently produce plaques in the absence of the polycation, substantiates the use of DEAE-dextran to perform a more reliable, faster and reproducible plaque assay of recombinant baculoviruses.

摘要

在通过空斑试验滴定野生型或重组杆状病毒时,在琼脂糖覆盖物中加入二乙氨基乙基葡聚糖(DEAE - 葡聚糖),会使病毒空斑的清晰度和对比度更高,并在感染后第5天至少将空斑数量和平均直径提高2倍。这种增加既与感染性病毒产量的增加无关,也与重组蛋白产量的增加无关,并且当聚阳离子仅存在于病毒接种物中或昆虫细胞单层与聚阳离子预孵育60分钟时不会发生。将这些观察结果扩展到多种不同来源的重组病毒,包括几种在没有聚阳离子的情况下不能持续产生空斑的杆状病毒,证实了使用DEAE - 葡聚糖对重组杆状病毒进行更可靠、更快且可重复的空斑试验。

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