Polyethylene glycol-mediated infection of non-permissive mammalian cells with semliki forest virus: application to signal transduction studies

被引:14
作者
Arudchandran, R
Brown, MJ
Song, JS
Wank, SA
Haleem-Smith, H
Rivera, J
机构
[1] NIAMSD, Sect Chem Immunol, NIH, Bethesda, MD 20892 USA
[2] NCI, Expt Immunol Branch, NIH, Bethesda, MD 20892 USA
[3] NIDDKD, Digest Dis Branch, NIH, Bethesda, MD 20892 USA
关键词
Semliki forest virus; polyethylene glycol; green fluorescent protein; signal transduction; protein expression;
D O I
10.1016/S0022-1759(98)00161-6
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Semliki Forest Virus (SFV) vectors allow the subcloning of a gene of interest directly in the expression vector, thus avoiding the need to select and purify viral recombinants, making this viral expression system attractive over many others for mammalian protein expression. We now describe a novel and generally applicable method for infection of non-permissive mammalian cells with SFV, that greatly enhances the utility of this expression system. We demonstrate that the hygroscopic polymer poly (ethylene glycol), PEG, promotes the infectivity of cells by SFV under conditions that did not promote cell-cell fusion. We also found that the PEG-induced infection and expression of an exogenous protein (green fluorescent protein, GFP) did not elevate the basal tyrosine kinase activity, induce a stress-activated responses, or result in aberrant cell responses. Expression of GFP tagged-Vav, an activator of stress-activated protein kinase (SAPK/JNK), resulted in the expected induction of JNK activity and in the normal redistribution of Vav in response to engagement of the high affinity receptor for IgE (Fc epsilon RI). Thus, our findings that PEG allows the infection of non-permissive cells by SFV makes this system extremely attractive for expression of proteins in mammalian cells, and studies on signal transduction and cellular localization in immune and non-immune cells. (C) 1999 Elsevier Science B.V. All rights reserved.
引用
收藏
页码:197 / 208
页数:12
相关论文
共 24 条
[1]   EXPRESSION OF BACTERIAL BETA-GALACTOSIDASE IN ANIMAL-CELLS [J].
AN, GH ;
HIDAKA, K ;
SIMINOVITCH, L .
MOLECULAR AND CELLULAR BIOLOGY, 1982, 2 (12) :1628-1632
[2]   Glycoprotein gH of pseudorabies virus is essential for penetration and propagation in cell culture and in the nervous system of mice [J].
Babic, N ;
Klupp, BG ;
Makoschey, B ;
Karger, A ;
Flamand, A ;
Mettenleiter, TC .
JOURNAL OF GENERAL VIROLOGY, 1996, 77 :2277-2285
[3]   IGE-INDUCED HISTAMINE-RELEASE FROM RAT BASOPHILIC LEUKEMIA-CELL LINES - ISOLATION OF RELEASING AND NON-RELEASING CLONES [J].
BARSUMIAN, EL ;
ISERSKY, C ;
PETRINO, MG ;
SIRAGANIAN, RP .
EUROPEAN JOURNAL OF IMMUNOLOGY, 1981, 11 (04) :317-323
[4]   SEMLIKI FOREST VIRUS EXPRESSION SYSTEM - PRODUCTION OF CONDITIONALLY INFECTIOUS RECOMBINANT PARTICLES [J].
BERGLUND, P ;
SJOBERG, M ;
GAROFF, H ;
ATKINS, GJ ;
SHEAHAN, BJ ;
LILJESTROM, P .
BIO-TECHNOLOGY, 1993, 11 (08) :916-920
[5]   ESTABLISHMENT OF AN IMMATURE MAST-CELL LINE FROM A PATIENT WITH MAST-CELL LEUKEMIA [J].
BUTTERFIELD, JH ;
WEILER, D ;
DEWALD, G ;
GLEICH, GJ .
LEUKEMIA RESEARCH, 1988, 12 (04) :345-355
[6]  
Chancellor JVM, 1997, QUAL LIFE RES, V6, P159
[7]   Use of green fluorescent protein variants to monitor gene transfer and expression in mammalian cells [J].
Cheng, LZ ;
Fu, J ;
Tsukamoto, A ;
Hawley, RG .
NATURE BIOTECHNOLOGY, 1996, 14 (05) :606-609
[8]  
Ciccarone VC, 1998, METH MOLEC MED, V13, P237, DOI 10.1385/0-89603-485-2:237
[9]  
Collins TL, 1997, IMMUNOL TODAY, V18, P221
[10]   Human HPK1, a novel human hematopoietic progenitor kinase that activates the JNK/SAPK kinase cascade [J].
Hu, MCT ;
Qiu, WR ;
Wang, XP ;
Meyer, CF ;
Tan, TH .
GENES & DEVELOPMENT, 1996, 10 (18) :2251-2264