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Shrimp recombinant baculovirus

Hansam Cho, Na Hye Park, Yuyeon Jang, Yong-Dae Gwon, Yeondong Cho, Yoon-Ki Heo, Ki-Hoon Park, Hee-Jung Lee, Tae Jin Choi, Young Bong Kim
Despite large economic losses attributable to white spot syndrome virus (WSSV), an infectious pathogen of penaeid shrimp and other crustaceans worldwide, no efficient vaccines or antiviral agents to control the virus are available at present. Here, we designed and constructed baculovirus-based vaccines delivering genes encoding the WSSV envelope proteins, VP28 and VP19. To enhance the immunogenicity of the baculovirus-based vaccine, we fused a Salmonella typhimurium flagellin 2 (FL2) gene with VP28 or VP19 gene...
March 2017: Journal of Invertebrate Pathology
Yingli Shi, Jianhai Xiang, Guangzhou Zhou, Tetsuzan Benny Ron, Hsin-I Tong, Wen Kang, Si Sun, Yuanan Lu
A newly isolated Pacific white shrimp (Litopenaeus vannamei) beta-actin promoter SbaP and its derivative compact construct SbaP (ENX) have recently been demonstrated to promote ectopic gene expression in vitro and in vivo. To further explore the potential transduction application, this newly isolated shrimp promoter SbaP was comparatively tested with cytomegalovirus (CMV), simian virus 40 (SV40), polyhedrin (Polh), and white spot syndrome virus immediate early gene 1 (WSSV ie1) four constitutive promoters and a beta-actin promoter (TbaP) from tilapia fish to characterize its promoting function in eight different cell lines...
June 2016: Marine Biotechnology
Triwit Rattanarojpong, Suthiwat Khankaew, Pongsak Khunrae, Rapeepun Vanichviriyakit, Kanokwan Poomputsa
White spot syndrome virus (WSSV) is a major causative agent in shrimp farming. Consequently, RNAi technology is an effective strategy to prevent WSSV infection in shrimp especially dsRNA targeting to rr2 of WSSV. In an effort to develop dsRNA expression in shrimp for control of WSSV infection, we developed a recombinant baculovirus expressing recombinant VP28 as the gene delivery system to carry a gene encoding dsRNA specific to rr2 for triggering the RNAi process in shrimp. The results showed that the recombinant baculovirus harboring VP28 was able to express VP28 indicated by Western blot with polyclonal antibody specific to VP28...
July 10, 2016: Journal of Biotechnology
Jayesh Puthumana, Priyaja Prabhakaran, Rosamma Philip, I S Bright Singh
In an attempt of in vitro transformation, transfection mediated expression of Simian virus-40 (T) antigen (SV40-T) and transduction mediated expression of Adenovirus type 12 early region 1A (12S E1A) oncogene were performed in Penaeus monodon lymphoid cells. pSV3-neo vector encoding SV40-T oncogene and a recombinant baculovirus BacP2-12S E1A-GFP encoding 12S E1A oncogene under the control of hybrid promoters were used. Electroporation and lipofection mediated transformation of SV40-T in lymphoid cells confirmed the transgene expression by phenotypic variation and the expression of GFP in co-transfection experiment...
December 2015: Fish & Shellfish Immunology
Jayesh Puthumana, Rosamma Philip, I S Bright Singh
It has been realized that shrimp cell immortalization may not be accomplished without in vitro transformation by expressing immortalizing gene in cells. In this process, efficiency of transgene expression is confined to the ability of vectors to transmit gene of interests to the genome. Over the years, unavailability of such vectors has been hampering application of such a strategy in shrimp cells. We report the use of recombinant baculovirus mediated transduction using hybrid promoter system for transgene expression in lymphoid cells of Penaeus monodon...
August 2016: Cytotechnology
Yi-Ting Yang, Der-Yen Lee, Yongjie Wang, Jer-Ming Hu, Wen-Hsiung Li, Jiann-Horng Leu, Geen-Dong Chang, Huei-Mien Ke, Shin-Ting Kang, Shih-Shun Lin, Guang-Hsiung Kou, Chu-Fang Lo
BACKGROUND: Penaeus monodon nudivirus (PmNV) is the causative agent of spherical baculovirosis in shrimp (Penaeus monodon). This disease causes significant mortalities at the larval stage and early postlarval (PL) stage and may suppress growth and reduce survival and production in aquaculture. The nomenclature and classification status of PmNV has been changed several times due to morphological observation and phylogenetic analysis of its partial genome sequence. In this study, we therefore completed the genome sequence and constructed phylogenetic trees to clarify PmNV's taxonomic position...
2014: BMC Genomics
Chun-Yi Hsiao, Yen-Ling Song
Hemocytes play important roles in crustacean immune responses. Generation of new hemocytes (hematopoiesis) is thus necessary to maintain homeostasis which is vital to crustaceans. In vertebrates, certain cytokines have been demonstrated to regulate hematopoiesis and immune responses. In invertebrates, however, little is known about cytokines related to hematopoiesis. In the present study, we cloned an astakine molecule from hemocytic cDNA of tiger shrimp (Penaeus monodon) which was 1509 bp in length with a 5'-UTR of 143 bp, a coding region of 375 bp and a 3'-UTR of 991 bp...
January 2010: Fish & Shellfish Immunology
S Syed Musthaq, Selvaraj Madhan, A S Sahul Hameed, Jimmy Kwang
White spot syndrome virus (WSSV) is a large dsDNA virus responsible for white spot disease in shrimp and other crustaceans. VP28 is one of the major envelope proteins of WSSV and plays a crucial role in viral infection. In an effort to develop a vaccine against WSSV, we have constructed a recombinant baculovirus with an immediate early promoter 1 which expresses VP28 at an early stage of infection in insect cells. Baculovirus expressed rVP28 was able to maintain its structural and antigenic conformity as indicated by immunofluorescence assay and western blot analysis...
September 1, 2009: Virology
Miao Li, Yu-Fei Wang, Yu Wang, Hui Gao, Na Li, Yuan Sun, Bing-Bing Liang, Hua-Ji Qiu
Non-replicating baculovirus-mediated gene transfer into mammalian cells has been developed as a vaccine strategy against a number of diseases in several animal models. In the present study, the BacMam vector, a baculovirus pseudotyped with the glycoprotein from vesicular stomatitis virus, was used as a recombinant vector to express classical swine fever virus (CSFV) E2 protein under the control of the immediate early 1 (ie1) promoter from shrimp white spot syndrome virus. The E2 gene was efficiently expressed in both insect and mammalian cells...
August 15, 2009: Immunology Letters
Yongjie Wang, Johannes A Jehle
Nudiviruses (NVs) are a highly diverse group of large, circular dsDNA viruses pathogenic for invertebrates. They have rod-shaped and enveloped nucleocapsids, replicate in the nucleus of infected host cells, and possess interesting biological and molecular properties. The unassigned viral genus Nudivirus has been proposed for classification of nudiviruses. Currently, the nudiviruses comprise five different viruses: the palm rhinoceros beetle virus (Oryctes rhinoceros NV, OrNV), the Hz-1 virus (Heliothis zea NV-1, HzNV-1), the cricket virus (Gryllus bimaculatus NV, GbNV), the corn earworm moth Hz-2 virus (HzNV-2), and the occluded shrimp Monodon Baculovirus reassigned as Penaeus monodon NV (PmNV)...
July 2009: Journal of Invertebrate Pathology
Yu Mi Ha, Yun Im Kim, Ki Hong Kim, Sung Koo Kim
This study was carried out to neutralize the WSSV one of the most virulent pathogen causing large economic damage in shrimp culture industry using the antiserum produced against recombinant WSSV envelope protein VP19 (rVP19) as a tool to evaluate WSSV infection mechanism. A fragment of VP19 was expressed in Sf21 insect cell using baculovirus expression system as fusion protein with 6 His-tag. Then, polyclonal antiserum against rVP19 was raised in white rabbit. A constant amount of WSSV (at 10(4) diluted stock) was incubated with various antiserum concentrations and injected into shrimp, Penaeus chinensis, for the neutralization challenge...
July 2008: Journal of Environmental Biology
Parin Chaivisuthangkura, Chutithorn Tawilert, Thanawan Tejangkura, Sombat Rukpratanporn, Siwaporn Longyant, Weerawan Sithigorngul, Paisarn Sithigorngul
The full-length of the occlusion body (OB) protein gene of Penaeus monodon nucleopolyhedrovirus (PemoNPV) was successfully isolated. The OB gene sequence contained an open reading frame (ORF) of 1359 nucleotides encoding a protein of 452 amino acid residues with a predicted molecular mass of 50.6 kDa. A putative late promoter element, TAAG, was identified 72 nt upstream of the translation start site. The amino acid sequences of tryptic digested peptides of PemoNPV OB protein obtained from LC-MS analysis matched quite well with various regions of deduced amino acid sequences...
November 25, 2008: Virology
Yu-Mi Ha, Gong Soo-Jung, Nguyen Thi-Hoai, Chae Hun Ra, Ki-Hong Kim, Yoon-Kwon Nam, Sung-Koo Kim
Two structural protein genes, VP19 and VP466, of white spot syndrome virus (WSSV) were cloned and expressed in Sf21 insect cells using a baculovirus expression system for the development of injection and oral feeding vaccines against WSSV for shrimps. The cumulative mortalities of the shrimps vaccinated by the injection of rVP19 and rVP466 at 15 days after the challenge with WSSV were 50.2% and 51.8%, respectively. For the vaccination by oral feeding of rVP19 and rVP466, the cumulative mortalities were 49.2% and 89...
May 2008: Journal of Microbiology and Biotechnology
Yu Wang, Hui Gao, Chuan Chen, Heping Zhao, Miao Li, Yuan Sun, Huaji Qiu
To compare the activity of different promoter in baculovirus-insect system, a series of recombinant baculoviruses were generated harboring the E-GFP reporter gene under the control of one of 5 promoters, including the ie1 promoter of shrimp white spot syndrome virus (WSSV), the truncated ie1 (mie1) promoter, the ETL promoter of the baculovirus, the elongated ETL (mETL) promoter, and the polyhedron promoter (P(PH)) of the baculovirus. The expression efficiency of the E-GFP reporter gene in the recombinant baculovirus-infected Sf9 cells was determined by flow cytometry...
April 2008: Sheng Wu Gong Cheng Xue Bao, Chinese Journal of Biotechnology
Siwaporn Longyant, Piengjan Poyoi, Parin Chaivisuthangkura, Thanawan Tejangkura, Weerawan Sithigorngul, Paisarn Sithigorngul, Sombat Rukpratanporn
The gene sequence encoding VP3 capsid protein of Taura syndrome virus (TSV) was cloned into pGEX-6P-1 expression vector and transformed into Escherichia coli BL21. After induction, recombinant GST-VP3 (rVP3) fusion protein was obtained and further purified by electro-elution before use in immunizing Swiss mice for production of monoclonal antibodies (MAb). One MAb specific to glutathione-S-transferase (GST) and 6 MAb specific to VP3 were selected using dot blotting and Western blotting. MAb specific to VP3 could be used to detect natural TSV infections in farmed whiteleg shrimp Penaeus vannamei by dot blotting and Western blotting, without cross reaction to shrimp tissues or other shrimp viruses, such as white spot syndrome virus (WSSV), yellow head virus (YHV), monodon baculovirus (MBV) and hepatopancreatic parvovirus (HPV)...
March 3, 2008: Diseases of Aquatic Organisms
Sonomi Hikima, Jun ichi Hikima, Jiraporn Rojtinnakorn, Ikuo Hirono, Takashi Aoki
Lysozyme cDNA was isolated from a kuruma shrimp, Marsupenaeus japonicus, hemocyte cDNA library. The cDNA consists of 1055 base pairs (bp) and encodes a chicken-type (c-type) lysozyme with a deduced amino acid sequence of 156 residues. The kuruma shrimp lysozyme has a high identity (79.7%) with pacific white shrimp lysozyme, and low to moderate identities (33.3-43.0%) with lysozymes of insects and vertebrates. Comparisons with other c-type lysozymes from invertebrates and vertebrates showed that the two catalytic residues (Glu58 and Asp75) and the eight cysteine residue motif were completely conserved...
October 16, 2003: Gene
Shinn-Tsuen Lin, Yun-Shiang Chang, Han-Ching Wang, Huey-Fen Tzeng, Zee-Fen Chang, Jung-Yaw Lin, Chung-Hsiung Wang, Chu-Fang Lo, Guang-Hsiung Kou
Infection of shrimp cells with white spot syndrome virus (WSSV) results in an increase in ribonucleotide reductase (RR) expression at the RNA level. In this article we further express and characterize the induction of a novel ribonucleotide reductase after WSSV infection of shrimp cells. A baculovirus/insect system was used to express the two recombinant protein subunits RR1 and RR2, and a DNA polymerase coupled RR activity assay showed a marked increase in ribonucleotide reductase activity when cell extracts containing recombinant RR1 and RR2 were combined...
December 20, 2002: Virology
Huey-Fen Tzeng, Zee-Fen Chang, Shao-En Peng, Chung-Hsiung Wang, Jung-Yaw Lin, Guang-Hsiung Kou, Chu-Fang Lo
The unique chimeric organization of the white spot syndrome virus (WSSV) tk-tmk gene encodes a protein which has significant homology to both cellular-type thymidine kinase (TK) and cellular-type thymidylate kinase (TMK), but the functional activity of this protein has not been demonstrated. Because TK is usually expressed only at very low levels in host cells, in this study, the coding region of WSSV tk-tmk was expressed in an insect/baculovirus expression system. The His-tagged recombinant WSSV TK-TMK was purified by affinity chromatography, and its enzyme activity was characterized by steady-state kinetics...
August 1, 2002: Virology
Li-Li Chen, Jiann-Horng Leu, Chang-Jen Huang, Chih-Ming Chou, Su-Mei Chen, Chung-Hsiung Wang, Chu-Fang Lo, Guang-Hsiung Kou
To identify the protein encoded by a 687-bp open reading frame (ORF) of a salI genomic DNA fragment of shrimp white spot syndrome virus (WSSV), we expressed the ORF in a baculovirus/insect cell expression system. The apparent molecular mass of the recombinant protein on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was 35 kDa in insect cells. Antibody raised against bacterially synthesized protein of the ORF identified a nucleocapsid protein (VP35) in the extracts of both the purified WSSV virions and the nucleocapsids which comigrated with the 35-kDa baculovirus-expressed recombinant protein on SDS-PAGE...
February 1, 2002: Virology
M C van Hulten, J Witteveldt, M Snippe, J M Vlak
White spot syndrome virus (WSSV) is a large DNA virus infecting shrimp and other crustaceans. The virus particles contain at least five major virion proteins, of which three (VP26, VP24, and VP15) are present in the rod-shaped nucleocapsid and two (VP28 and VP19) reside in the envelope. The mode of entry and systemic infection of WSSV in the black tiger shrimp, Penaeus monodon, and the role of these proteins in these processes are not known. A specific polyclonal antibody was generated against the major envelope protein VP28 using a baculovirus expression vector system...
July 5, 2001: Virology
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