scholarly journals Production of vaccines for treatment of infectious diseases by transgenic plants

2016 ◽  
Vol 107 (1) ◽  
pp. 191 ◽  
Author(s):  
Kristina LEDL ◽  
Zlata LUTHAR
Keyword(s):  
Transgenic Plants ◽  
Infectious Diseases ◽  
Disease Virus ◽  
Hepatitis B Surface Antigen ◽  
Expression System ◽  
Expression Vectors ◽  
Norwalk Virus ◽  
Mouth Disease Virus ◽  
Avian Influenza Virus H5n1 ◽  
Transformation Methods

<p>Since the first pathogen antigen was expressed in transgenic plants with the aim of producing edible vaccine in<em> </em>early 1990s, transgenic plants have become a well-established expression system for production of alternative vaccines against various human and animal infectious diseases. The main focus of plant expression systems in the last five years has been on improving expression of well-studied antigens such as porcine reproductive and respiratory syndrome (PRRSV), bovine viral diarrhea disease virus (BVDV), footh and mouth disease virus (FMDV), hepatitis B surface antigen (HBsAg), rabies G protein, rotavirus, Newcastle disease virus (NDV), Norwalk virus capsid protein (NVCP), avian influenza virus H5N1, <em>Escherichia coli</em> heat-labile enterotoxin subunit B (LT-B), cholera toxin B (CT-B), human immunodeficiency virus (HIV), artherosclerosis, ebola and anthrax. Significant increases in expression have been obtained using improved expression vectors, different plant species and transformation methods.</p>

scholarly journals Construction of CRISPR/Cas9 expression vectors habouring gRNA targeted on SLIAA9 gene of tomato

2020 ◽  
Vol 18 (1) ◽  
pp. 147-156
Author(s):  
Bui Manh Minh ◽  
Ha Hong Hanh ◽  
Le Thi Thu Hien ◽  
Huynh Thi Thu Hue
Keyword(s):  
Transgenic Plants ◽  
Genome Editing ◽  
Tomato Fruit ◽  
Expression System ◽  
Secondary Compounds ◽  
Expression Vectors ◽  
Tomato Plants ◽  
Normal Morphology ◽  
Transgenic Tomato Plants ◽  
Pcr Test

Tomato (Solanum lycopersicum) is a nutritious fruit containing many secondary compounds with health benefits. The formation of tomato fruit through fertilization is controlled by auxin through Aux/IAA9 and ARF8 proteins. The mutated SlIAA9 gene leads to the parthenocarpic development of fruit or seedless tomato fruit. Nowadays, the CRISPR/Cas9 genome editing system is becoming increasingly popular in modifying desired genes on plant objects. In this study, gRNAs which target on tomato SlIAA9 gene were designed and inserted into CRISPR/Cas9 vectors. In addition, two strains of A. tumefaciens harboring pRGEB31-IAA9G2 and pRGEB32-IAA9G2 vectors carrying CRISPR/Cas9 expression system towards SlIAA9 gene in tomato were successfully created. The strain of A. tumefaciens harboring pRGEB31- IAA9G2 plasmid was used to develop transgenic tomato plants from Micro-Tom variety. PCR test showed that 5/14 plants had the presence of Cas9 gene in T0 plants. The transgenic plants have a normal morphology in comparation with the controls. The evaluation of mutant efficiency, type, and stability of mutations on the SlIAA9 will be conducted on next-generation plants when the mutations are stable and segregated into descendents.

scholarly journals Assembly and characterization of foot-and-mouth disease virus empty capsid particles expressed within mammalian cells

2013 ◽  
Vol 94 (8) ◽  
pp. 1769-1779 ◽  
Author(s):  
Maria Gullberg ◽  
Bartosz Muszynski ◽  
Lindsey J. Organtini ◽  
Robert E. Ashley ◽  
Susan L. Hafenstein ◽  
...  
Keyword(s):  
Disease Virus ◽  
Mammalian Cells ◽  
Expression System ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Capsid Proteins ◽  
Dependent Manner ◽  
Empty Capsid ◽  
Mouth Disease Virus ◽  
Foot And Mouth

The foot-and-mouth disease virus (FMDV) structural protein precursor, P1-2A, is cleaved by the virus-encoded 3C protease (3Cpro) into the capsid proteins VP0, VP1 and VP3 (and 2A). In some systems, it is difficult to produce large amounts of these processed capsid proteins since 3Cpro can be toxic for cells. The expression level of 3Cpro activity has now been reduced relative to the P1-2A, and the effect on the yield of processed capsid proteins and their assembly into empty capsid particles within mammalian cells has been determined. Using a vaccinia-virus-based transient expression system, P1-2A (from serotypes O and A) and 3Cpro were expressed from monocistronic cDNA cassettes as P1-2A-3C, or from dicistronic cassettes with the 3Cpro expression dependent on a mutant FMDV internal ribosome entry site (IRES) (designated P1-2A-mIRES-3C). The effects of using a mutant 3Cpro with reduced catalytic activity or using two different mutant IRES elements (the wt GNRA tetraloop sequence GCGA converted, in the cDNA, to GAGA or GTTA) were analysed. For both serotypes, the P1-2A-mIRES-3C construct containing the inefficient GTTA mutant IRES produced the highest amount of processed capsid proteins. These products self-assembled to form FMDV empty capsid particles, which have a related, but distinct, morphology (as determined by electron microscopy and reconstruction) from that determined previously by X-ray crystallography. The assembled empty capsids bind, in a divalent cation-dependent manner, to the RGD-dependent integrin αvβ6, a cellular receptor for FMDV, and are recognized appropriately in serotype-specific antigen ELISAs.

scholarly journals Evidence for the role of His-142 of protein 1C in the acid-induced disassembly of foot-and-mouth disease virus capsids

1999 ◽  
Vol 80 (8) ◽  
pp. 1911-1918 ◽  
Author(s):  
Fiona M. Ellard ◽  
Jeff Drew ◽  
Wendy E. Blakemore ◽  
David I. Stuart ◽  
Andrew M. Q. King
Keyword(s):  
Disease Virus ◽  
Expression System ◽  
Foot And Mouth Disease ◽  
Dipole Interaction ◽  
Virus Genome ◽  
Mouth Disease ◽  
Mouth Disease Virus ◽  
Foot And Mouth ◽  
Acidic Conditions ◽  
Α Helix

Foot-and-mouth disease virus (FMDV) capsids are inherently labile under mildly acidic conditions, dissociating to pentamers at pH values in the region of 6·5, with the release of protein 1A and the viral RNA. This acid-induced disassembly is thought to be required for the entry of the virus genome into the host cell. Previous work has highlighted a histidine–α-helix charge-dipole interaction at the twofold axes of symmetry between pentamers and has suggested that this interaction plays a role in acid-induced disassembly. The validity of this theory has now been tested by converting the implicated residue, His-142 of protein 1C, to Arg, Phe and Asp. The effects of such changes were studied by using a previously described vaccinia virus expression system, in which synthesis and processing of FMDV capsid proteins results in the self-assembly of capsids. In agreement with the histidine–α-helix charge-dipole theory, assembly in the arginine mutant was found to be greatly reduced, while capsids of the aspartic acid mutant were considerably more stable under acidic conditions than the wild-type. Aberrant but acid-stable complexes were obtained in the phenylalanine mutant.

A novel methodology to develop a foot and mouth disease virus (FMDV) peptide-based vaccine in transgenic plants

Vaccine ◽  
2002 ◽  
Vol 20 (7-8) ◽  
pp. 1141-1147 ◽  
Author(s):  
Marı́a J Dus Santos ◽  
Andrés Wigdorovitz ◽  
Karina Trono ◽  
Raúl D Rı́os ◽  
Pascual M Franzone ◽  
...  
Keyword(s):  
Transgenic Plants ◽  
Disease Virus ◽  
Foot And Mouth Disease ◽  
Mouth Disease ◽  
Mouth Disease Virus ◽  
Foot And Mouth

scholarly journals Expression of erythropoietin in Indian tetraploid potato variety

F1000Research ◽  
2012 ◽  
Vol 1 ◽  
pp. 26
Author(s):  
Priti N Desai ◽  
Harish Padh
Keyword(s):  
Transgenic Plants ◽  
Mammalian Cells ◽  
Capital Investment ◽  
Transgene Expression ◽  
Present Report ◽  
Expression System ◽  
Expression Vectors ◽  
Pcr Analysis ◽  
Tissue Specific ◽  
Tissue Specific Promoter

With the advent of protein-based biotech drugs in the market, the quest for the “perfect” protein expression system, which is both economical and effective, has come into focus. Currently bacteria, yeast, insect cells, mammalian cells, transgenic animal and transgenic plants are widely used for the expression of therapeutic proteins. Among these, transgenic plants provide advantages in terms of low production cost, lower capital investment in infrastructure, and suitable post-translational modifications. The major limitation of plants as an expression host is the low level of transgene expression. To increase the expression of heterologous proteins in plants, a number of approaches have been used. One of the approaches is to increase the transgene expression by using tissue-specific promoter(s) which can concentrate the protein of interest in targeted tissues and, thus, prove advantageous in downstream purification. In the present report, a protocol for expression of heterologous protein erythropoietin in potato tuber using patatin, the tuber-tissue-specific promoter, was standardized. Expression vectors for production of the erythropoietin gene under tissue-specific promoter were successfully constructed. For production of a transgenic plant, tissue culture techniques for regeneration of the whole plant from single explants were standardized. Polymerase chain reaction (PCR) analysis was performed to confirm the stable integration of the erythropoietin gene in the potato plant by using sequence-specific primers.

scholarly journals EXPRESSION AND PURIFICATION CAPSID PROTEINS VP0, VP1 AND VP3 OF FOOT AND MOUTH DISEASE VIRUS TYPE O IN ESCHERICHIA COLI

2016 ◽  
Vol 54 (5) ◽  
pp. 597
Author(s):  
Nguyen Hoang Duong ◽  
Chi-Ning Chuang ◽  
Nguyen Phuong Hoa ◽  
Tran Thi Kim Dung ◽  
Le Hong Minh ◽  
...  
Keyword(s):  
Disease Virus ◽  
Expression System ◽  
Foot And Mouth Disease ◽  
Initial Step ◽  
Mouth Disease ◽  
Capsid Proteins ◽  
Virus Like Particles ◽  
Safety Issues ◽  
Mouth Disease Virus ◽  
Foot And Mouth

Foot and mouth disease virus (FMDV) causing infectious disease affects broadly cloven-hoofed animals. It has 7 different serotypes. However O is the most prevalent type in 3 founded types (O, A, Asia1) recurrence in Vietnam fields. The current vaccine for FMDV is inactivated or attenuated forms. Vaccines were able to raise strong immune responses but still caused the safety concerns. Virus-like particles could be a new vaccine generation that fulfills the present questions. On the one hand, it can tackle the safety issues, on the other hand, it can reserve FMDV intrinsic form which will provoke high immunogenicity. The important initial step to make virus-like particles is expression the capsid proteins of FMDV in appropriate system. So this study we describe how to design, express and purify of all capsid proteins: VP0, VP1 and VP3 of O type FMDV isolated in Vietnam field using SUMO fusion expression system.

Sign in / Sign up

Export Citation Format

Share Document