Browse > Article
http://dx.doi.org/10.7852/ijie.2011.23.1.147

Expression of Pseudorabies Virus (PRV) Glycoproteins gB, gC and gD using Bacterial Expression System  

Yun, Bit-Na-Rae (Department of Agricultural Biology, College of Agriculture, Life & Environment Science, Chungbuk National University)
Bae, Sung-Min (Department of Agricultural Biology, College of Agriculture, Life & Environment Science, Chungbuk National University)
Lee, Jun-Beom (Department of Agricultural Biology, College of Agriculture, Life & Environment Science, Chungbuk National University)
Kim, Hee-Jung (Department of Agricultural Biology, College of Agriculture, Life & Environment Science, Chungbuk National University)
Woo, Soo-Dong (Department of Agricultural Biology, College of Agriculture, Life & Environment Science, Chungbuk National University)
Publication Information
International Journal of Industrial Entomology and Biomaterials / v.23, no.1, 2011 , pp. 147-153 More about this Journal
Abstract
The Pseudorabies (PR), also called Aujeszky's disease (AD), is an infectious viral disease caused by an alpha herpes virus and has domestic and wild pigs, as well as a wide range of domestic and wild animals, as the natural host. Pseudorabies virus (PRV) virions contain several envelope glycoproteins. Among them, gB, gC and gD are regarded as the major immunogenic proteins. We expressed these glycoproteins using the bacterial expression system and analyzed recombinant proteins. Expression of glycoproteins gC and gD were observed on SDS-PAGE or Western blot analysis, but gB was not. Optimal concentration of IPTG and inducing time were determined as 1.0 mM and 4 h, respectively, for the expression of both gC and gD in E. coli. A sodium dodecyl sulfate (SDS) was the most efficient detergent in solubilizing insoluble recombinant protein.
Keywords
Pseudorabies virus; Glycoproteins gB, gC and gD; Bacterial expression system;
Citations & Related Records
Times Cited By KSCI : 1  (Citation Analysis)
연도 인용수 순위
1 Rabhi-Essafi I, Sadok A, Khalaf N, Fathallah DM (2007) A strategy for high-level expression of soluble and functional human interferon a as a GST-fusion protein in E.coli. Protein Eng Des Sel 20(5), 201-209.   DOI   ScienceOn
2 Semba H, Ichige E, Imanaka T, Atomi H, Aoyagi H (2008) Efficient production of active form of recombinant cassava hydroxynitrilelyase using Escherichia coli in low-temperature culture. Appl Microbiol Biotechnol 79(4), 563-569.   DOI   ScienceOn
3 Singh SM, Panda AK (2005) Solubilization and refolding of bacterial inclusion body proteins. J Biosci Bioeng 99(4), 303-310.   DOI   ScienceOn
4 Song HS, Lee SH, Oh EH, Park TH (2009) Expression, solubilization and purification of a human olfactory receptor from Escherichia coli. Curr Microbiol 59(3), 309-314.   DOI   ScienceOn
5 Sorensen HP, Mortensen KK (2005) Advanced genetic strategies for recombinant protein expression in Escherichia coli. J Biotechnol 115(2), 113-128.   DOI   ScienceOn
6 Thakur MK, Ghosh S (2010) GST-tagged mouse estrogen receptor alpha-transactivation domain fusion protein is specifically degraded during its over-expression in E. coli and purification. Mol Biol Rep 37(3), 1335-1440.   DOI   ScienceOn
7 Vilnis A, Sussman MD, Thacker BJ, Senn M, Maes RK (1998) Vaccine genotype and route of administration affect pseudorabies field virus latency load after challenge. Vet Microbiol 62(2), 81-96.   DOI   ScienceOn
8 Bane SE, Velasquez JE, Robinson AS (2007) Expression and purification of milligram levels of inactive G-protein coupled receptors in E. coli. Protein Expr Purif 52(2), 348-355.   DOI   ScienceOn
9 Assadi-Porter FM, Patry S, Markley JL (2008) Efficient and rapid protein expression and purification of small high disulfide containing sweet protein brazzein in E. coli. Protein Expr Purif 58(2), 263-268.   DOI   ScienceOn
10 Balasch M, Pujols J, Segalés J, Pumarola M (1998) Aujeszky's disease (pseudorabies) virus detection in cerebrospinal fluid in experimentally infected pigs. Vet Microbiol 60(2-4), 99- 106.   DOI   ScienceOn
11 Burgess-Brown NA, Sharma S, Sobott F, Loenarz C, Oppermann U, Gileadi O (2008) Codon optimization can improve expression of human genes in Escherichia coli: A multigene study. Protein Expr Purif 59(1), 94-102.   DOI   ScienceOn
12 Katzen F, Peterson TC, Kudlicki W (2009) Membrane protein expression: no cells required. Trends Biotechnol 27(8), 455- 460.   DOI   ScienceOn
13 Grabowska AK, Lipiñska AD, Rohde J, Szewczyk B, Bienkowska- Szewczyk K, Rziha HJ (2009) New baculovirus recombinants expressing Pseudorabies virus (PRV) glycoproteins protect mice against lethal challenge infection. Vaccine 27(27), 3584-3591.   DOI   ScienceOn
14 Hong W, Xiao S, Zhou R, Fang L, He Q, Wu B, Zhou F, Chen H (2002) Protection induced by intramuscular immunization with DNA vaccines of pseudorabies in mice, rabbits and piglets. Vaccine 20, 1205-1214.   DOI   ScienceOn
15 Hsu FS, Lee RC (1984) Use of hyperimmune serum, vaccination, and certain management procedures for control of pseudorabies in swine. J Am Vet Med Assoc 184, 1463-1466.
16 Liu YJ, Song FP, He KL, Yuan Y, Zhang XX, Gao P, Wang JH, Wang GY (2004) Expression of a modified Cry1Ie gene in E. coli and in transgenic tobacco confers resistance to corn borer. Acta Biochim Biophys Sin (Shanghai) 36(4), 309-313.   DOI   ScienceOn
17 Kirschner A, Altenbuchner J, Bornscheuer UT (2007) Cloning, expression, and characterization of a Baeyer-Villigermonooxygenase from Pseudomonas fluorescens DSM 50106 in E. coli. Appl Microbiol Biotechnol 73(5), 1065-1072.
18 Klupp BG, Hengartner CJ, Mettenleiter TC, Enquist LW (2004) Complete, Annotated Sequence of the Pseudorabies Virus Genome. J Virol 78, 424-440.   DOI   ScienceOn
19 Koo HN, Bae SM, Choi JB, Shin TY, Yun BNR, Choi JY, Lee KS, Roh JY, Je YH, Jin BR, Yoo SS, Kim JS, Kim YI, Yoon IJ, Woo SD (2011) Characterization and expression of the pseudorabies virus (NYJ strain) glycoproteins in Bombyxmori cells and larvae. J Asia-Pacific Entomol 14, 107-117.   DOI   ScienceOn
20 Le PU, Lenferink AE, Pinard M, Baardsnes J, Massie B, O'Connor-McCourt MD (2009) Escherichia coli expression and refolding of E/K-coil-tagged EGF generates fully bioactive EGF for diverse applications. Protein Expr Purif 64(2), 108-117.   DOI   ScienceOn
21 Mettenleiter TC (1994) Pseudorabies (Aujeszky's disease) virus: state of the art. Acta Vet Hung 42(2-3), 153-177.
22 Ong RM, Goh KM, Mahadi NM, Hassan O, Rahman RN, Illias RM (2008) Cloning, extracellular expression and characterization of a predominant beta-CGTase from Bacillus sp. G1 in E. coli. J Ind Microbiol Biotechnol 35(12), 1705-1714.   DOI   ScienceOn
23 Mettenleiter TC (1996) Immunobiology of pseudorabies (Aujeszky's disease). Vet Immunol Immunopathol 54(1-4), 221-229.   DOI   ScienceOn
24 Neidhardt EA, Punreddy SR, McLean JE, Hedstrom L, Grossman TH (1999) Expression and characterization of E. coliproduced soluble, functional human dihydroorotate dehydrogenase: a potential target for immunosuppression. J MolMicrobiol Biotechnol 1(1), 183-188.