排序方式: 共有35条查询结果,搜索用时 15 毫秒
11.
A genetically encoded photosensitizer 总被引:2,自引:0,他引:2
Bulina ME Chudakov DM Britanova OV Yanushevich YG Staroverov DB Chepurnykh TV Merzlyak EM Shkrob MA Lukyanov S Lukyanov KA 《Nature biotechnology》2006,24(1):95-99
Photosensitizers are chromophores that generate reactive oxygen species (ROS) upon light irradiation. They are used for inactivation of specific proteins by chromophore-assisted light inactivation (CALI) and for light-induced cell killing in photodynamic therapy. Here we report a genetically encoded photosensitizer, which we call KillerRed, developed from the hydrozoan chromoprotein anm2CP, a homolog of green fluorescent protein (GFP). KillerRed generates ROS upon irradiation with green light. Whereas known photosensitizers must be added to living systems exogenously, KillerRed is fully genetically encoded. We demonstrate the utility of KillerRed for light-induced killing of Escherichia coli and eukaryotic cells and for inactivating fusions to beta-galactosidase and phospholipase Cdelta1 pleckstrin homology domain. 相似文献
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Guliĭ OI Zaĭtsev BD Kuznetsova IE Shikhabudinov AM Karavaeva OA Dykman LA Staroverov SA Ignatov OV 《Biofizika》2012,57(3):460-467
The phage mini-antibodies to bacterial cells of strain Azospirillum brasilense Sp245 were obtained and the possibility of using them for detection of microbial cells by means of a lateral field excited piezoelectric resonator was studied. It has been found that the frequency dependencies of the real and imaginary parts of the electrical impedance of the resonator loaded by the cell suspension A. brasilense Sp245 with the mini-antibodies, significantly differ from those of the resonator with the control cell suspension without mini-antibodies. The concentration limit of possible determination of the microbial cells in their interaction with the mini-antibodies is equal to 10(3) cells/ml. It has been ascertained that detection of A. brasilense Sp245 cells using the mini-antibodies is possible even in the presence of other cultures, for example, E. coli BL-Ril and A. brasilense Sp7 cells. Therefore, it has been shown for the first time that detection of microbial cells by an electro-acoustic sensor is feasible. 相似文献
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Computer analysis predicted a strong donor splice site within the 3'-part of the far-red fluorescent protein Katushka coding region. To test the functional activity of this site a model vector has been constructed. This vector encoded Katushka and green fluorescent protein TagGFP2 with a gene fragment of tafazzin in between. Normal splicing of this pre-mRNA should result in a frameshift between Katushka and TagGFP2. Alternatively, after splicing at internal katushka donor splice site appearance of Katushka-TagGFP2 fusion protein was expected. Expression of this construct in a mammalian cell culture led to bright red and green fluorescence. Therefore, katushka-specific donor splice site is functional. Disruption of this splice site by several silent substitutions resulted in red-only fluorescent cells that corresponded to normal splicing. The mutant katushka can be used for visualization of pre-mRNA splicing at single cell level by fluorescence microscopy and flow cytometry. 相似文献
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The mammalian pannexin family is homologous to the invertebrate innexin gap junction proteins 总被引:8,自引:0,他引:8
Baranova A Ivanov D Petrash N Pestova A Skoblov M Kelmanson I Shagin D Nazarenko S Geraymovych E Litvin O Tiunova A Born TL Usman N Staroverov D Lukyanov S Panchin Y 《Genomics》2004,83(4):706-716
We have cloned the genes PANX1, PANX2 and PANX3, encoding putative gap junction proteins homologous to invertebrate innexins, which constitute a new family of mammalian proteins called pannexins. Phylogenetic analysis revealed that pannexins are highly conserved in worms, mollusks, insects and mammals, pointing to their important function. Both innexins and pannexins are predicted to have four transmembrane regions, two extracellular loops, one intracellular loop and intracellular N and C termini. Both the human and mouse genomes contain three pannexin-encoding genes. Mammalian pannexins PANX1 and PANX3 are closely related, with PANX2 more distant. The human and mouse pannexin-1 mRNAs are ubiquitously, although disproportionately, expressed in normal tissues. Human PANX2 is a brain-specific gene; its mouse orthologue, Panx2, is also expressed in certain cell types in developing brain. In silico evaluation of Panx3 expression predicts gene expression in osteoblasts and synovial fibroblasts. The apparent conservation of pannexins between species merits further investigation. 相似文献
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D. V. Pristensky S. A. Staroverov D. N. Ermilov S. Yu. Shchyogolev L. A. Dykman 《Biochemistry (Moscow) Supplemental Series B: Biomedical Chemistry》2007,1(3):249-253
Penetration of ivermectin adsorbed on the surface of colloidal gold particles or included in micelles to phagocytic cells of the immune system was studied. Detection of the preparation in the cells was carried out by the immunochemical method, employing miniantibodies to ivermectin obtained from a combinatory phage library and by the chromatographic method using the HPLC system “Stayer.” It was found that accumulation of ivermectin adsorbed on colloidal gold particles was accumulated in the cells most intensively than in the case of other carriers. 相似文献
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Yu. G. Yanushevich N. G. Gurskaya D. B. Staroverov S. A. Lukyanov K. A. Lukyanov 《Russian Journal of Bioorganic Chemistry》2003,29(4):325-329
The gene of a new red fluorescent protein zoan2RFP from coral polyp Zoanthus sp., a homologue of the known green fluorescent protein from the jellyfish Aequorea victoria, was cloned. At early stages of maturation, zoan2RFP exhibits green fluorescence, which then turns to the red one. A similar phenomenon was recently reported for the E5 mutant of the red fluorescent coral protein DsRed. Zoan2RFP differs from E5 by faster maturation kinetics and the complete disappearance of green fluorescence in the mature protein. Naturally occurring proteins of this type can be considered as intermediate forms between the green and red fluorescent proteins, which are formed during the microevolution of fluorescent proteins. 相似文献
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O. I. Guliy B. D. Zaitsev A. M. Shikhabudinov A. A. Teplykh I. A. Borodina S. A. Pavliy O. S. Larionova A. S. Fomin S. A. Staroverov L. A. Dykman O. V. Ignatov 《Applied Biochemistry and Microbiology》2016,52(4):457-463
It has been demonstrated that electroacoustic analysis with polyclonal antibodies can be used for bacteriophage detection. The frequency dependences of the real and imaginary parts of electrical impedance of a resonator with a viral suspension with antibodies were shown to be essentially different from the dependences of a resonator with control viral suspension without antibodies. It was shown that ΦAl-Sp59b bacteriophages were detected with the use of antibodies in the presence of foreign virus particles. The ΦAl-Sp59b bacteriophage content in the analyzed suspension was ~1010–106 phages/mL; the time of analysis was no more than 5 min. The optimally informative parameter for obtaining reliable information was the change in the real or imaginary part of electrical impedance at a fixed frequency near the resonance upon the addition of specific antibodies to the analyzed suspension. It was demonstrated that the interaction between bacteriophages and antibodies can be recorded, offering good prospects for the development of a biological sensor for liquid-phase identification and virus detection. 相似文献
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Anisimova VE Rebrikov DV Zhulidov PA Staroverov DB Lukyanov SA Shcheglov AS 《Biochemistry. Biokhimii?a》2006,71(5):513-519
We overexpressed duplex-specific nuclease (DSN) from Kamchatka crab in Escherichia coli cells and developed procedures for purification, renaturation, and activation of this protein. We demonstrated identity of the properties of the native and recombinant DSN. We also successfully applied the recombinant DSN for full-length cDNA library normalization. 相似文献