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Nagesh K. Tripathi Jyoti Shukla Karttik C. Biswal P. V. Lakshmana Rao 《Applied microbiology and biotechnology》2010,86(6):1795-1803
Japanese encephalitis (JE) is one of the leading causes of acute encephalopathy affecting children and adolescents in the
tropics. Optimization of media was carried out for enhanced production of recombinant JE virus envelope domain III (EDIII)
protein in Escherichia coli. Furthermore, batch and fed-batch cultivation process in E. coli was also developed in optimized medium. Expression of this protein in E. coli was induced with 1 mM isopropyl-β-thiogalactoside and yielded an insoluble protein aggregating to form inclusion bodies.
The inclusion bodies were solubilized in 8 M urea, and the protein was purified under denaturing conditions using Ni-NTA affinity
chromatography. After fed-batch cultivation, the recombinant E. coli resulted in cell dry weight and purified protein about 36.45 g l−1 and 720 mg l−1 of culture, respectively. The purity of the recombinant JE virus EDIII protein was checked by sodium dodecyl sulfate polyacrylamide
gel electrophoresis analysis, and reactivity of this protein was determined by Western blotting and ELISA with JE virus-infected
human serum samples. These results establish the application of this protein to be used for the diagnosis of JE virus infection
or for further studies in vaccine development. This process may also be suitable for the high-yield production of other recombinant
viral proteins. 相似文献
94.
R. K. Sharma M. S. Negi S. Sharma P. Bhardwaj R. Kumar E. Bhattachrya S. B. Tripathi D. Vijayan A. R. Baruah S. C. Das B. Bera R. Rajkumar J. Thomas R. K. Sud N. Muraleedharan M. Hazarika M. Lakshmikumaran S. N. Raina P. S. Ahuja 《Biochemical genetics》2010,48(7-8):549-564
India has a large repository of important tea accessions and, therefore, plays a major role in improving production and quality of tea across the world. Using seven AFLP primer combinations, we analyzed 123 commercially important tea accessions representing major populations in India. The overall genetic similarity recorded was 51%. No significant differences were recorded in average genetic similarity among tea populations cultivated in various geographic regions (northwest 0.60, northeast and south both 0.59). UPGMA cluster analysis grouped the tea accessions according to geographic locations, with a bias toward China or Assam/Cambod types. Cluster analysis results were congruent with principal component analysis. Further, analysis of molecular variance detected a high level of genetic variation (85%) within and limited genetic variation (15%) among the populations, suggesting their origin from a similar genetic pool. 相似文献
95.
Sanjay Dwivedi R. D. Tripathi Sudhakar Srivastava Ragini Singh Amit Kumar Preeti Tripathi Richa Dave U. N. Rai Debasis Chakrabarty P. K. Trivedi R. Tuli B. Adhikari M. K. Bag 《Protoplasma》2010,245(1-4):113-124
The exposure of paddy fields to arsenic (As) through groundwater irrigation is a serious concern that may not only lead to As accumulation to unacceptable levels but also interfere with mineral nutrients in rice grains. In the present field study, profiling of the mineral nutrients (iron (Fe), phosphorous, zinc, and selenium (Se)) was done in various rice genotypes with respect to As accumulation. A significant genotypic variation was observed in elemental retention on root Fe plaque and their accumulation in various plant parts including grains, specific As uptake (29–167 mg kg?1 dw), as well as As transfer factor (4–45%). Grains retained the least level of As (0.7–3%) with inorganic As species being the dominant forms, while organic As species, viz., dimethylarsinic acid and monomethylarsonic acid, were non-detectable. In all tested varieties, the level of Se was low (0.05–0.12 mg kg?1 dw), whereas that of As was high (0.4–1.68 mg kg?1 dw), considering their safe/recommended daily intake limits, which may not warrant their human consumption. Hence, their utilization may increase the risk of arsenicosis, when grown in As-contaminated areas. 相似文献
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Sanjay K Singh Vinayak R Tripathi Rakesh K Jain Surendra Vikram Satyendra K Garg 《Microbial cell factories》2010,9(1):59
Background
Many workers have reported halotolerant bacteria from saline conditions capable of protease production. However, antibiotic resistance and heavy metal tolerance pattern of such organisms is not documented very well. Similarly, only a few researchers have reported the pattern of pH change of fermentation medium during the course of protease production. In this study, we have isolated a halotolerant Bacillus cereus SIU1 strain from a non-saline environment and studied its antibiotic and heavy metal resistance pattern. The isolate produces a thermoalkaline protease and changes the medium pH during the course of fermentation. Thermostability of protease was also studied for 30 min. 相似文献98.
Wild type p53 exists in a constant state of equilibrium between wild type and mutant conformation and undergoes conformational changes at elevated temperature. We have demonstrated that the co-chaperone CHIP (carboxyl terminus of Hsp70-interacting protein), which suppressed aggregation of several misfolded substrates and induced the proteasomal degradation of both wild type and mutant p53, physically interacts with the amino terminus of WT53 and prevented it from irreversible thermal inactivation. CHIP preferentially binds to the p53 mutant phenotype and restored the DNA binding activity of heat-denatured p53 in an ATP-independent manner. In cells under elevated temperatures that contained a higher level of p53 mutant phenotype, CHIP restored the native-like conformation of p53 in the presence of geldanamycin, whereas CHIP-small interfering RNA considerably increased the mutant form. Further, under elevated temperatures, the levels of CHIP and p53 were higher in nucleus, and chromatin immunoprecipitation shows the presence of p53 and CHIP together upon the DNA binding site in the p21 and p53 promoters. We propose that CHIP might be a direct chaperone of wild type p53 that helps p53 in maintaining wild type conformation under physiological condition as well as help resurrect p53 mutant phenotype into a folded native state under stress condition. 相似文献
99.
In our search for synthetic substitutes for artemisinin and its derivatives we had earlier prepared a series of adamantane-based 1, 2, 4-trioxanes 5a-c which had shown promising activity against P. berghei in Swiss mice. We have further evaluated these and two new compounds (5d-e) against Plasmodium knowlesi W1, a virulent malaria parasite in rhesus monkeys, in the dose range of 40-80 mg/kg x 5 days by intramuscular route. Trioxanes 5b and 5c showed 100% protection and cure at 80 mg/kg x 5 days, while trioxane 5a showed 71% cure at this dose. Detailed studies again showed 50% curative effect of 5a at 40 mg/kg x 5 days treatment. 相似文献
100.