Publications (16)36.25 Total impact
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Dataset: Localization of Rabies Virus Glycoprotein into the Endoplasmic
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Article: Receptors of garlic (Allium sativum) lectins and their role in insecticidal action.
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ABSTRACT: Garlic (Allium sativum) lectins are promising candidate molecules for the protection against chewing (lepidopteran) as well as sap sucking (homopteran) insect pests. Molecular mechanism of toxicity and interaction of lectins with midgut receptor proteins has been described in many reports. Lectins show its effect right from sensory receptors of mouth parts by disrupting the membrane integrity and food detection ability. Subsequently, enter into the gut lumen and interact with midgut glycosylated proteins like alkaline phosphatase (ALP), aminopeptidase-N (APN), cadherin-like proteins, polycalins, sucrase, symbionin and others. These proteins play critical role in life cycle of insect directly or indirectly. Lectins interfere with the activity of these proteins and causes physiological disorders leading to the death of insects. Lectins further transported across the insect gut, accumulated in various body parts (like haemolymph and ovary) and interact with intracellular proteins like symbionin and cytochrome p450. Binding with cytochrome p450 (which involve in ecdysone synthesis) might interfere in the development of insects, which results in growth retardation and pre-mature death.The Protein Journal 05/2012; 31(6):439-46. · 1.04 Impact Factor -
Article: Localization of rabies virus glycoprotein into the endoplasmic reticulum produces immunoprotective antigen.
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ABSTRACT: Rabies virus surface glycoprotein (rabies G-protein) with (G+RS) and without (G-RS) endoplasmic reticulum retrieval signal was expressed and characterized in tobacco plants. Transgenically expressed rabies G-protein was estimated at 0.015-0.38 % of total leaf protein. The relative migration of the rabies G-protein on SDS-PAGE was at the position, as anticipated for the viral coat protein (~66 kDa). Immunolocalization by confocal microscopy established that immunoprotective G+RS expressed in tobacco was primarily confined to ER. G+RS showed binding to Con A lectin and was susceptible to N-glycosidase F activity similar to native rabies G-protein. However, the G-RS transgenically expressed in tobacco leaves was glycosylated differently and was resitant to N-glycosidase F. Immunological studies and Rapid Fluorescent Foci Inhibition Test (RFFIT) showed that G+RS was immunogenic and immunoprotective, whereas G-RS was moderately immunogenic but non-protective against live virus challenge. Hence, plants can express the antigenic component of rabies virus with suitable glycosylation, which is important to give protection against rabies virus infection.The Protein Journal 05/2012; 31(6):447-56. · 1.04 Impact Factor -
Article: Enhanced expression of rabies virus surface G-protein in Escherichia coli using SUMO fusion.
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ABSTRACT: Fusion systems are known to increase the expression of difficult to express recombinant proteins in soluble form to facilitate their purification. Rabies glycoprotein was also tough to express at sufficient level in soluble form in both E. coli and plant. The present work was aimed to over-express and purify this membrane protein from soluble extract of E. coli. Fusion of Small Ubiqutin like Modifier (SUMO) with rabies glycoprotein increased ~1.5 fold higher expression and ~3.0 fold solubility in comparison to non-fused in E. coli. The SUMO fusion also simplified the purification process. Previously engineered rabies glycoprotein gene in tobacco plants provides complete protection to mice, but the expression was very low for purification. Our finding demonstrated that the SUMO-fusion was useful for enhancing expression and solubility of the membrane protein and again proves to be a good alternative technology for applications in biomedical and pharmaceutical research.The Protein Journal 12/2011; 31(1):68-74. · 1.04 Impact Factor -
Article: Role of alkaline phosphatase in insecticidal action of Cry1Ac against Helicoverpa armigera larvae.
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ABSTRACT: Cry1Ac δ-endotoxin produced by Bacillus thuringiensis (Bt) is used as a bio-pesticide for the control of Helicoverpa armigera. Aminopeptidases N (APN) and alkaline phosphatase (ALP) play critical roles in its action against H. armigera larvae. The binding of Cry1Ac with brush border membrane vesicle (BBMV) proteins was increased with the larval development although the sensitivity of larvae to δ-endotoxins decreased. There was higher expression of ALP than APN in early instar larvae with a ~10-fold higher affinity of Cry1Ac towards ALP than to APN. Binding to a specific receptor is therefore more important for the insecticidal activity rather than overall binding to the BBMV proteins. ALP might play a major role in toxicity as compared to APN.Biotechnology Letters 06/2011; 33(10):2027-36. · 1.68 Impact Factor -
Patent: Chimeric G-protein based rabies vaccine.
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ABSTRACT: A novel chimeric protein of rabies virus designed to express a chimeric G protein at a high level in transgenic plants. A gene was also designed and chemically synthesized to encode the chimeric G protein and expressed at high level in plant tissue. The gene was expressed in transgenic tobacco plants to examine its therapeutic efficacy against infection by rabies virus. The chimeric G protein was enriched in plant membranes. The BalbC mice were immunized with the plant leaf expressed G protein. Plant derived G protein elicited higher immune response as compared to the commercial vaccine. The mice displayed protective immunity when they were challenged with live virus. Chimeric G protein expressed at high level in plants leaves was demonstrated to function as a commercially valuable subunit vaccine against rabies vaccine infection.Ref. No: US7901691B2, Year: 03/2011 -
Article: BECLIN1 from Arabidopsis thaliana under the generic control of regulated expression systems, a strategy for developing male sterile plants.
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ABSTRACT: Induction of male sterility followed by successful outcrossing is a prerequisite for hybrid seed production. In this article, we have identified a novel use of the BECLIN 1 gene of Arabidopsis, in inducing male sterility in plants, when expressed in the anther tapetum of tobacco. We also report a stringently regulated and high-level expression of the desired gene in tapetum by using a two-component transcription regulation system. The tapetum-specific, two-component transcription system utilizes the TGTA-TBPm³ complementation principle that has been demonstrated by us earlier. We also report a glucocorticoid-dependent expression of AtBECLIN 1 in tapetum, thereby developing glucocorticoid-inducible male sterility in plants.Plant Biotechnology Journal 12/2010; 8(9):1005-22. · 5.44 Impact Factor -
Article: Expression of δ-endotoxin Cry1EC from an inducible promoter confers insect protection in peanut (Arachis hypogaea L.) plants.
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ABSTRACT: Spodoptera litura (F.) is a polyphagous foliage insect and a major pest on peanut (Arachis hypogaea L.). Constitutive expression of δ-endotoxin Cry1EC gives protection against S. litura, as reported earlier. In this study, insect bites and salicylic acid induced high-level expression of Cry1EC was achieved in peanut. In order to achieve this, the expression of pathogenesis responsive promoter PR-1a was enhanced by placing it downstream of the CaMV35S promoter in the pCAMBIA 1300 backbone. The resultant promoter CaMV35S(r)PR-1a expressed a high level of insecticidal δ-endotoxin Cry1EC. The Gus expression under the control of CaMV35S(r)PR-1a served as a convenient marker for evaluation of promoter response to different treatments. Transgenic events that showed a very low level of uninduced expression and no expression in seeds were selected. The Cry1EC expression in leaves increased nearly eightfold in the selected event, following induction by salicylic acid. Both the salicylic-acid-treated and the S. litura-bitten leaves showed the highest expression after 2 days. Leaves from salicylic-acid-induced transgenic plants caused 100% mortality of S. litura at all stages of larval development. The results suggest that high expression of inducible promoters provides a good strategy for the development of safer transgenic food and feed crops.Pest Management Science 10/2010; 67(2):137-45. · 2.25 Impact Factor -
Article: Expression of δ-endotoxin Cry1EC from an inducible promoter confers insect protection inExpression of δ-endotoxin Cry1EC from an inducible promoter confers insect protection in peanut peanut
Pest Management Science 01/2010; 67::137–145. · 2.25 Impact Factor -
Article: Universal plant DNA barcode loci may not work in complex groups: a case study with Indian berberis species.
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ABSTRACT: The concept of DNA barcoding for species identification has gained considerable momentum in animals because of fairly successful species identification using cytochrome oxidase I (COI). In plants, matK and rbcL have been proposed as standard barcodes. However, barcoding in complex genera is a challenging task. We investigated the species discriminatory power of four reportedly most promising plant DNA barcoding loci (one from nuclear genome--ITS, and three from plastid genome--trnH-psbA, rbcL and matK) in species of Indian Berberis L. (Berberidaceae) and two other genera, Ficus L. (Moraceae) and Gossypium L. (Malvaceae). Berberis species were delineated using morphological characters. These characters resulted in a well resolved species tree. Applying both nucleotide distance and nucleotide character-based approaches, we found that none of the loci, either singly or in combinations, could discriminate the species of Berberis. ITS resolved all the tested species of Ficus and Gossypium and trnH-psbA resolved 82% of the tested species in Ficus. The highly regarded matK and rbcL could not resolve all the species. Finally, we employed amplified fragment length polymorphism test in species of Berberis to determine their relationships. Using ten primer pair combinations in AFLP, the data demonstrated incomplete species resolution. Further, AFLP analysis showed that there was a tendency of the Berberis accessions to cluster according to their geographic origin rather than species affiliation. We reconfirm the earlier reports that the concept of universal barcode in plants may not work in a number of genera. Our results also suggest that the matK and rbcL, recommended as universal barcode loci for plants, may not work in all the genera of land plants. Morphological, geographical and molecular data analyses of Indian species of Berberis suggest probable reticulate evolution and thus barcode markers may not work in this case.PLoS ONE 01/2010; 5(10):e13674. · 4.09 Impact Factor -
Article: Rabies glycoprotein fused with B subunit of cholera toxin expressed in tobacco plants folds into biologically active pentameric protein.
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ABSTRACT: The pentameric B subunit of cholera toxin (CtxB) is an efficient mucosal adjuvant for vaccines. We report the expression of a chimeric protein comprising the synthetic cholera toxin B subunit fused at its C-terminal with rabies surface glycoprotein (G protein) in tobacco plants. The approximately 80.3 kDa fusion polypeptide expressed at 0.4% of the total soluble protein in leaves of the selected transgenic lines. The fusion protein formed a approximately 403 kDa pentameric protein which was functionally active in binding to GM1 receptor. The plant-made protein had a higher affinity for GM1 receptor than the native bacterial CtxB. The pentameric fusion protein was recognized by the anti-cholera toxin as well as anti-rabies antibodies. Its immuno-protective ability against rabies remains to be examined.Protein Expression and Purification 10/2009; 70(2):184-90. · 1.59 Impact Factor -
Article: Efficient production of gossypol from hairy root cultures of cotton (Gossypium hirsutum L.).
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ABSTRACT: A protocol for induction and establishment of Agrobacterium rhizogenes mediated hairy root culture of Gossypium hirsutum was developed through infection with the A4 strain and co-cultivation on hormone-free semi-solid MS medium with B5 vitamins. It resulted in the emergence of hairy roots from the leaf explants, 21 days after infection. The transformation of hairy roots was established by PCR amplification of rol B and rol C genes of the Ri plasmid. All root lines expressed gossypol, although distinct inter-clonal quantitative variations were noticed. Five independent hairy root lines were studied for their growth kinetics as well as gossypol production. The yield potentials of one of them superseded others, as well as the non-transformed, in-vitro grown control roots. The content of gossypol in hairy roots reached a level of 2.43 mg/g DW. It was 4.5 times higher than in vitro and 1.47 times higher than in vivo grown roots of G. hirsutum. Selection of high gossypol producing hairy root lines of G. hirsutum can provide an alternative source for large-scale production of gossypol.Current pharmaceutical biotechnology 09/2009; 10(7):691-700. · 3.40 Impact Factor -
Article: Plants as bioreactors for the production of vaccine antigens.
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ABSTRACT: Plants have been identified as promising expression systems for commercial production of vaccine antigens. In phase I clinical trials several plant-derived vaccine antigens have been found to be safe and induce sufficiently high immune response. Thus, transgenic plants, including edible plant parts are suggested as excellent alternatives for the production of vaccines and economic scale-up through cultivation. Improved understanding of plant molecular biology and consequent refinement in the genetic engineering techniques have led to designing approaches for high level expression of vaccine antigens in plants. During the last decade, several efficient plant-based expression systems have been examined and more than 100 recombinant proteins including plant-derived vaccine antigens have been expressed in different plant tissues. Estimates suggest that it may become possible to obtain antigen sufficient for vaccinating millions of individuals from one acre crop by expressing the antigen in seeds of an edible legume, like peanut or soybean. In the near future, a plethora of protein products, developed through 'naturalized bioreactors' may reach market. Efforts for further improvements in these technologies need to be directed mainly towards validation and applicability of plant-based standardized mucosal and edible vaccines, regulatory pharmacology, formulations and the development of commercially viable GLP protocols. This article reviews the current status of developments in the area of use of plants for the development of vaccine antigens.Biotechnology advances 05/2009; 27(4):449-67. · 8.25 Impact Factor -
Article: Conserved nucleotide sequences in highly expressed genes in plants
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ABSTRACT: Genes that code for proteins expressed at high and low levels in plants were classified into separate data sets. The two data sets were analysed to identify the conserved nucleotide sequences that may characterize genes with contrasting levels of expression. The AUG context that characterized the highly expressed genes is (A/C)N2AAN3(A/T)T(A/C) AACAATGGCTNCC(T/A)CNA(C/T)(A/C). The data set of highly expressed genes shows overrepresentation of codons for alanine at the second position and serine at the third and fourth positions after the translation initiation codon. The characteristic transcription initiation site in the highly expressed genes is CAN(A/C)(A/C)(C/A)C(C/A)N2A(C/A). The promoter region is characterized by two tandemly repeated TATA elements, sometimes with one and rarely with two point mutations in the highly expressed genes. Besides the two tandemly repeated TATA elements, the promoter context in the highly expressed genes is overrepresented by C, C and G at the -3, -1 and+9 positions respectively. The characteristic TATA motif in the highly expressed plant genes is (T/C)(T/A)N2TCACTATATATAG. Most of these features are not present in the genes ubiquitously expressed at low levels in plants.Journal of Genetics 04/1999; 78(2):123-131. · 1.09 Impact Factor -
Article: In vitro proliferation of shoots and regeneration of cotton
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ABSTRACT: Shoot proliferation from different explants of several Indian cultivars of cotton was studied in culture. Cotyledonary nodes taken along with the shoot apex of seedlings produced multiple shoots on modified MS nutrient agar supplemented with cytokinins. 6-Benzyladenine was most effective in inducing growth of multiple shoots. Explants of several genotypes formed organogenic masses that differentiated to secondary shoots on repeated subculture. The isolated shoots were rooted on basal medium supplemented with naphthaleneacetic acid and were transferred to soil after acclimatization.Plant Cell Tissue and Organ Culture 10/1997; 51(2):149-152. · 3.09 Impact Factor -
Article: The extent of genetic diversity among Vanilla species: Comparative results for RAPD and ISSR
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ABSTRACT: Vanilla is a large genus of about 110 species in the orchid family (Orchidaceae), including the species Vanilla planifolia from which commercial vanilla flavoring is derived. Since most species of vanilla are considered rare and endangered there is an urgent need to conserve them through genetic analysis and propagation/conservation studies on this crop.The present study investigated the genetic diversity among nine leafy- and leaf-less Vanilla species employing 30 decamer RAPD primers and 10 ISSR primers. The species under study were diverse and displayed a range of variability (0–66% and 0–81% for RAPD and ISSR, respectively). A total of 154 RAPD polymorphic markers (83.24%, h = 0.378) and 93 ISSR polymorphic markers (86.11%, h = 0.363) were used to generate a genetic similarity matrix followed by the cluster analysis. Specific groupings were revealed by each cluster analysis with slight variation between two different markers. Among the nine species studied, V. planifolia, Vanilla aphylla and Vanilla tahitensis revealed very low level of variation within their collections, thus indicating a narrow genetic base. The large genetic distance of Vanilla andamanica from other species suggests its different origin. A close genetic affinity was observed between the pairs V. planifolia, V. tahitensis and Vanilla albida, V. aphylla. These are the first comparative results for RAPD and ISSR reporting inter-relationship among nine cultivated, wild and hybrid Vanilla species.Industrial Crops and Products.
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Institutions
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1997–2012
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National Botanical Research Institute
Lucknow, Uttar Pradesh, India
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2010
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National Agri-Food Biotechnology Institute
Mohali, State of Punjab, India
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1999
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National Botanical Research Institute - India
Lucknow, Uttar Pradesh, India
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