Expression and secretion of recombinant outer-surface protein A from the lyme disease agent, Borrelia burgdorferi, in Nicotiana tabacum suspension cells

被引：10

作者：

Navarre, Catherine ^{[1
]}

Delannoy, Melanie ^{[1
]}

Lefebvre, Benoit ^{[1
]}

Nader, Joseph ^{[1
]}

Vanham, Delphine ^{[1
]}

Boutry, Marc ^{[1
]}

机构：

[1] Catholic Univ Louvain, Inst Sci Vie, Unite Biochim Physiol, B-1348 Louvain, Belgium

来源：

TRANSGENIC RESEARCH | 2006年 / 15卷 / 03期

关键词：

glycosylation; heterologous expression; OspA; plant; suspension cells;

D O I：

10.1007/s11248-006-0002-7

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

The ospA gene of Borrelia burgdorferi codes for an outer membrane lipoprotein, which is a major antigen of the Lyme disease agent. Recombinant OspA vaccines tested so far were expressed in Escherichia coli. In this study, we investigated the expression of a soluble OspA protein in Nicotiana tabacum suspension cells and evaluated the secretion of OspA driven by either its own bacterial signal peptide or a plant signal peptide fused to the amino-terminal cysteine of the mature form. In both cases, the signal peptide was cleaved off and OspA secreted. During secretion, OspA was N-glycosylated. Addition of a C-terminal KDEL sequence led to retention of OspA in the endoplasmic reticulum.

引用

页码：325 / 335

页数：11

共 37 条

[1] HIGH-EFFICIENCY TRANSFORMATION OF CULTURED TOBACCO CELLS [J].

AN, GH .

PLANT PHYSIOLOGY, 1985, 79 (02) :568-570

[2] A Rab1 GTPase is required for transport between the endoplasmic reticulum and Golgi apparatus and for normal Golgi movement in plants [J].

Batoko, H ;

Zheng, HQ ;

Hawes, C ;

Moore, I .

PLANT CELL, 2000, 12 (11) :2201-2217

[3] Never say never again: protein glycosylation in pathogenic bacteria [J].

Benz, I ;

Schmidt, MA .

MOLECULAR MICROBIOLOGY, 2002, 45 (02) :267-276

[4] Analysis of the lipidated recombinant outer surface protein A from Borrelia burgdorferi by mass spectrometry [J].

Bouchon, B ;

Klein, M ;

Bischoff, R ;

VanDorsselaer, A ;

Roitsch, C .

ANALYTICAL BIOCHEMISTRY, 1997, 246 (01) :52-61

[5]

BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3

[6] Saturation of the endoplasmic reticulum retention machinery reveals anterograde bulk flow [J].

Crofts, AJ ;

Leborgne-Castel, N ;

Hillmer, S ;

Robinson, DG ;

Phillipson, B ;

Carlsson, LE ;

Ashford, DA ;

Denecke, J .

PLANT CELL, 1999, 11 (11) :2233-2247

[7]

Dunn J J, 1990, Protein Expr Purif, V1, P159, DOI 10.1016/1046-5928(90)90011-M

[8] ROLE OF ATTACHED LIPID IN IMMUNOGENICITY OF BORRELIA-BURGDORFERI OSPA [J].

ERDILE, LF ;

BRANDT, MA ;

WARAKOMSKI, DJ ;

WESTRACK, GJ ;

SADZIENE, A ;

BARBOUR, AG ;

MAYS, JP .

INFECTION AND IMMUNITY, 1993, 61 (01) :81-90

[9] LONG-TERM PROTECTION OF MICE FROM LYME-DISEASE BY VACCINATION WITH OSPA [J].

FIKRIG, E ;

BARTHOLD, SW ;

KANTOR, FS ;

FLAVELL, RA .

INFECTION AND IMMUNITY, 1992, 60 (03) :773-777

[10] Reactivity with a specific epitope of outer surface protein A predicts protection from infection with the Lyme disease spirochete, Borrelia burgdorferi [J].

Golde, WT ;

Piesman, J ;

Dolan, MC ;

Kramer, M ;

Hauser, P ;

Lobet, Y ;

Capiau, C ;

Desmons, P ;

Voet, P ;

Dearwester, D ;

Frantz, JC .

INFECTION AND IMMUNITY, 1997, 65 (03) :882-889

← 1 2 3 4 →