A method of alphavirus replicon particle titration based on expression of functional replicase/transcriptase

被引：2

作者：

Gangolli, SS ^{[1
]}

Vasilakis, N ^{[1
]}

Kovacs, GR ^{[1
]}

Zamb, TJ ^{[1
]}

Kowalski, J ^{[1
]}

机构：

[1] Wyeth Vaccines, Viral Vaccine Discovery Res, Pearl River, NY 10965 USA

来源：

JOURNAL OF VIROLOGICAL METHODS | 2003年 / 109卷 / 02期

关键词：

alphaviruses; Venezuelan equine encephalitis virus; replicon particles; titration;

D O I：

10.1016/S0166-0934(03)00063-6

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Alphavirus replicon particles are being exploited for a variety of purposes both in vitro as gene expression vectors, and in vivo as vaccines or gene therapy vectors. There is a need for a simple and universal method of. titration of replicon particles that is independent of expression of the foreign protein. We devised a method that uses modified vaccinia virus Ankara (MVA) as an indicator virus, to deliver a Venezuelan equine encephalitis virus (VEE) defective helper RNA encoding green fluorescent protein (GFP). Co-infection of cells with the MVA-based indicator and Venezuelan equine encephalitis virus replicon particles (VRP) results in expression of the GFP gene. VRP titer is readily determined by counting fluorescent cells. (C) 2003 Elsevier Science B.V. All rights reserved.

引用

页码：133 / 138

页数：6

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