Expression of an RSV-gag virus-like particle in insect cell lines and silkworm larvae

被引：28

作者：

Deo, Vipin Kumar ^{[1
]}

Tsuji, Yoshitaka ^{[2
]}

Yasuda, Tomomi ^{[2
]}

Kato, Tatsuya ^{[2
]}

Sakamoto, Naonori ^{[3
]}

Suzuki, Hisao ^{[4
]}

Park, Enoch Y. ^{[1
,2
]}

机构：

[1] Shizuoka Univ, Grad Sch Sci & Technol, Biotechnol Lab, Integrated Biosci Sect,Suruga Ku, Shizuoka 4228529, Japan

[2] Shizuoka Univ, Dept Appl Biol Chem, Biotechnol Lab, Fac Agr, Shizuoka 4228529, Japan

[3] Shizuoka Univ, Dept Mat Sci & Chem Engn, Naka Ku, Shizuoka 4328561, Japan

[4] Shizuoka Univ, Grad Sch Sci & Technol, Naka Ku, Shizuoka 4328561, Japan

来源：

JOURNAL OF VIROLOGICAL METHODS | 2011年 / 177卷 / 02期

关键词：

Rous sarcoma virus group antigen protein; Virus like particle; Bombyx mori nucleopolyhedrovirus bacmid; Silkworm; Insect cell; ROUS-SARCOMA-VIRUS; ASSEMBLED IN-VITRO; P10; DOMAIN; HUMAN BETA-1,3-N-ACETYLGLUCOSAMINYLTRANSFERASE; PROTEIN; DELIVERY; MICROSCOPY; MORPHOLOGY; SYSTEM; SIZE;

D O I：

10.1016/j.jviromet.2011.07.012

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Rous sarcoma virus group antigen protein-based virus-like particles (VLPs) are well known for their structural integrity and ease of handling. VLPs play an important role in drug delivery systems because they can be manipulated with ease. In this study, a new method was established for expressing Rous sarcoma virus group antigen protein based VLPs in silkworm larvae and establishing stably expressing insect cell lines. These VLPs have been isolated by ultracentrifugation using a sucrose step gradient of 10-60% (v/v), and their spherical structure has been confirmed using transmission electron microscopy (TEM). The spherical morphology is similar in both the silkworm larvae and in stably expressing cell lines. Silkworm larvae are better suited for producing Rous sarcoma virus group antigen protein-based VLPs on a large scale; yields from silkworm larvae were approximately 8.2-fold higher than yields from stable cell lines. These VLPs provide a new method for large-scale application in vaccine development and drug delivery systems. (C) 2011 Elsevier B.V. All rights reserved.

引用

页码：147 / 152

页数：6

共 29 条

[1] The retroviral capsid domain dictates virion size, morphology, and coassembly of Gag into virus-like particles [J].

Ako-Adjei, D ;

Johnson, MC ;

Vogt, VM .

JOURNAL OF VIROLOGY, 2005, 79 (21) :13463-13472

[2] Cryo-electron microscopy reveals conserved and divergent features of Gag packing in immature particles of rous sarcoma virus and human immunodeficiency virus [J].

Briggs, JAG ;

Johnson, MC ;

Simon, MN ;

Fuller, SD ;

Vogt, VM .

JOURNAL OF MOLECULAR BIOLOGY, 2006, 355 (01) :157-168

[3] In vitro assembly of virus-like particles with Rous sarcoma virus Gag deletion mutants: Identification of the p10 domain as a morphological determinant in the formation of spherical particles [J].

Campbell, S ;

Vogt, VM .

JOURNAL OF VIROLOGY, 1997, 71 (06) :4425-4435

[4] Biochemical characterization of Rous sarcoma virus MA protein interaction with membranes [J].

Dalton, AK ;

Murray, PS ;

Murray, D ;

Vogt, VM .

JOURNAL OF VIROLOGY, 2005, 79 (10) :6227-6238

[5]

Deo VK, 2006, J VIROL METHODS, V134, P212, DOI 10.1016/j.jviromet.2006.01.007

[6] Multiple co-transfection and co-expression of human β-1,3-N-acetylglucosaminyltransferase with human calreticulin chaperone cDNA in a single step in insect cells [J].

Deo, VK ;

Park, EY .

BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, 2006, 43 (129-135) :129-135

[7] Production of scFv-displaying BmNPV in silkworm larvae and its efficient purification [J].

Dojima, Takashi ;

Nishina, Takuya ;

Kato, Tatsuya ;

Ueda, Hiroshi ;

Park, Enoch Y. .

BIOTECHNOLOGY AND APPLIED BIOCHEMISTRY, 2010, 57 :63-69

[8] Comparison of the N-linked glycosylation of human β1,3-N-acetylglucosaminyltransferase 2 expressed in insect cells and silkworm larvae [J].

Dojima, Takashi ;

Nishina, Takuya ;

Kato, Tatsuya ;

Uno, Btsuyoshi ;

Yagi, Hirokazu ;

Kato, Koichi ;

Park, Enoch Y. .

JOURNAL OF BIOTECHNOLOGY, 2009, 143 (01) :27-33

[9] Phylogenetically important regions of the influenza A H1 hemagglutinin protein [J].

Fanning, TG ;

Taubenberger, JK .

VIRUS RESEARCH, 1999, 65 (01) :33-42

[10] The C-terminal half of TSG101 blocks rous sarcoma virus budding and sequesters Gag into unique nonendosomal structures [J].

Johnson, MC ;

Spidel, JL ;

Ako-Adjei, D ;

Wills, JW ;

Vogt, VM .

JOURNAL OF VIROLOGY, 2005, 79 (06) :3775-3786

← 1 2 3 →