绿色荧光蛋白作为筛选重组杆状病毒的工具。

Green fluorescent protein as a tool for screening recombinant baculoviruses.

作者信息

Eriksson S, Raivio E, Kukkonen J P, Eriksson K, Lindqvist C

机构信息

Abo Akademi University, Department of Biochemistry and Pharmacy, Finland.

出版信息

J Virol Methods. 1996 May;59(1-2):127-33. doi: 10.1016/0166-0934(96)02032-0.

DOI:10.1016/0166-0934(96)02032-0

PMID:8793839

Abstract

The gene encoding the green fluorescent protein (GFP) from the jellyfish Aequorea victoria, ligated to the honeybee melittin signal peptide-encoding sequence, was inserted under transcriptional control of the polyhedrin promoter of the Autographa californica nuclear polyhedrosis virus and expressed in the Spodoptera frugiperda insect cell line Sf9 during viral infection. The recombinant green fluorescent protein was identified by SDS-PAGE gel electrophoresis followed by Coomassie blue staining of lysates from the recombinant baculovirus infected insect cells. Emission and excitation scanning of the recombinant baculovirus infected insect cells gave an emission maximum of 509 nm and excitation maximum of 398 nm. The GFP protein expressed was also detected in infected insect cells by a flow cytometer analysis.

摘要

将来自维多利亚多管水母的绿色荧光蛋白（GFP）编码基因与蜜蜂蜂毒肽信号肽编码序列连接，插入到苜蓿银纹夜蛾核型多角体病毒多角体蛋白启动子的转录控制下，并在病毒感染期间在草地贪夜蛾昆虫细胞系Sf9中表达。通过SDS-PAGE凝胶电泳，随后对重组杆状病毒感染的昆虫细胞裂解物进行考马斯亮蓝染色，鉴定重组绿色荧光蛋白。对重组杆状病毒感染的昆虫细胞进行发射和激发扫描，发射最大值为509 nm，激发最大值为398 nm。还通过流式细胞仪分析在感染的昆虫细胞中检测到表达的GFP蛋白。