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  1. 9 Acta Biochimica Polonica

  2. 3 BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology

  3. 2 Acta Parasitologica

  4. 2 Biotechnologia

  5. 2 Wiadomości Parazytologiczne. Suplement

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  1. 2 Dabrowski S.

  2. 2 Kur J.

  3. 2 Pawelczyk T.

  4. 2 Sakowicz M.

  5. 1 Aguado-Martinez A.

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  1. 1 2023

  2. 2 2021

  3. 1 2018

  4. 1 2015

  5. 1 2014

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1

Prokaryotic toxin-antitoxin systems - the role in bacterial physiology and application in molecular biology

100%
Bukowski M., Rojowska A., Wladyka B.
Acta Biochimica Polonica
|
2011
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tom 58
|
nr 1
 Bacteria have developed multiple complex mechanisms ensuring an adequate response to environmental changes. In this context, bacterial cell division and growth are subject to strict control to ensure metabolic balance and cell survival. A plethora of studies cast light on toxin-antitoxin (TA) systems as metabolism regulators acting in response to environmental stress conditions. Many of those studies suggest direct relations between the TA systems and the pathogenic potential or antibiotic resistance of relevant bacteria. Other studies point out that TA systems play a significant role in ensuring stability of mobile genetic material. The evolutionary origin and relations between various TA systems are still a subject of a debate. The impact of toxin-antitoxin systems on bacteria physiology prompted their application in molecular biology as tools allowing cloning of some hard-to-maintain genes, plasmid maintenance and production of recombinant proteins.
2

Antisense oligonucleotides to PAI-1 mRNA as potential agents for therapeutic intervention in cardiovascular diseases

100%
Cierniewski C. S., Pawlowska Z., Pluskota E., Stasiak M., Kobylanska A., Misiura K., Maciaszek A., Koziolkiewicz M., Stec W. J.
Biotechnologia
|
1996
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nr 4
3

Recombinant His-tagged DNA polymerase. II. Cloning and purification of Thermus aquaticus recombinant DNA polymerase [Stoffel fragment]

100%
Dabrowski S., Kur J.
Acta Biochimica Polonica
|
1998
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tom 45
|
nr 3
The Stoffel DNA fragment, shortened by 12 bp from 5' end, coding for StoffelDNA polymerase (missing 4 amino acids at N-terminus of Stoffel amino-acids sequence) from the thermophilic Thermus aquaticus (strain YT-1) was amplified, cloned and expressed in Escherichia coli. The recombinant Stoffel fragment contained a polyhistidine tag at the N-terminus (21 additional amino acids) that allowed its single-step isolation by Ni2+ affinity chromatography. The enzyme was characterized and displayed high DNA polymerase activity and thermostability evidently higher than the native Tag DNA polymerase.
4

Expression level of Ubc9 protein in rat tissues

100%
Golebiowski F., Szulc A., Sakowicz M., Szutowicz A., Pawelczyk T.
Acta Biochimica Polonica
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2003
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tom 50
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nr 4
Ubc9 is a homologue of the E2 ubiquitin conjugating enzyme and participates in the covalent linking of SUMO-1 molecule to the target protein. In this report we describe a simple and efficient method for obtaining pure human recombinant Ubc9 protein. The purified Ubc9 retained its native structure and was fully active in an in vitro sumoylation assay with the promyelocytic leukaemia (PML) peptide as a substrate. In order to better understand the physiology of Ubc9 protein we examined its levels in several rat tissues. Immunoblot analyses performed on tissue extracts revealed quantitative and qualitative differences in the expression pattern of Ubc9. The Ubc9 protein was present at a high level in spleen and lung. Moderate level of Ubc9 was detected in kidney and liver. Low amount of Ubc9 was observed in brain, whereas the 18 kDaband of Ubc9 was barely visible or absent in heart and skeletal muscle. In heart and muscle extracts the Ubc9 antibodies recognized a 38 kDa protein band. This band was not visible in extracts of other rat tissues. A comparison of the relative levels of Ubc9 mRNA and protein indicated that the overall expression level of Ubc9 was the highest in spleen and lung. In spleen, lung, kidney, brain, liver and heart there was a good correlation between the 18 kDa protein and Ubc9 mRNA levels. In skeletal muscle the Ubc9 mRNA level was unproportionally high comparing to the level of the 18 kDa protein. The presented data indicate that in the rat the expression of the Ubc9 protein ap­pears to have some degree of tissue specificity.
5

Recombinant His-tagged DNA polymerase. I. Cloning, purification and partial characterization of Thermus thermophilus recombinant DNA polymerase

100%
Dabrowski S., Kur J.
Acta Biochimica Polonica
|
1998
|
tom 45
|
nr 3
The Tth DNA polymerase gene from the thermophilic Thermus thermophilus (strain HB8) was amplified, cloned and expressed in Escherichia coli. The recombinant DNA polymerase containing a polyhistidine tag at the N-terminus was isolated in a single step by Ni2+ affinity chromatography. The purified recombinant enzyme, showing high polymerase activity contained 43 additional amino-acid residues (including a cluster of six histidine residues inserted for purification of the recombinant protein by metal-affinity chromatography) at N-terminus. The applied overexpression system was very efficient giving 700000 u of DNA polymerase activity from 1 liter of induced culture. The enzyme was characterized and displayed high DNA polymerase and reverse transcriptase activities and high thermostability as compared to the native Tht DNA polymerase.
6

Concanavalin A-agarose removes mannan impurities from an extracellularly expressed Pichia pastoris recombinant protein

100%
Palczewska M., Batta G., Groves P.
Cellular and Molecular Biology Letters
|
2003
|
tom 08
|
nr 3
Pichia pastoris secretes few native proteins. However, the more than 1 g 1-1 of extracellularly expressed mannan interfered with the purification of our extracellularly expressed, non-glycosylated recombinant protein. Concanavalin A-agarose removed more than 95% of the unwanted mannan as monitored by phenol reaction. A 13C-based NMR assay confirmed this improvement. Concanavalin A-agarose can assist the purification of extracellular expressed, non-glycosylated proteins from yeasts.
7

Using combined recombinant protein in the diagnosis of bovine brucellosis

84%
Bulashev A., Jakubowski T., Mukantayev K., Tursunov K., Kiyan V., Zhumalin A.
Medycyna Weterynaryjna
|
2018
|
tom 74
|
nr 03
The aim of this research was to obtain a combined recombinant protein consisting of immunodominant regions of Brucella outer membrane proteins (Omps) with the molecular weights of 25 kDa (Omp25) and 31 kDa (Omp31). The search for candidate proteins was carried out using NCBI PubMed and NCBI GenBank databases. The two most immunodominant regions of Omps were selected. The first region was in a position from 48 to 83 amino acids of Omp31, while the second one was in the position from 180 to 224 amino acids of Omp25. This combined sequence was designated as OmpBm-Ba. The pET32 vector was used for cloning and the expression of the combined recombinant protein in E. coli BL21(DE3). The antigenicity of recombinant OmpBm-Ba (rOmpBm-Ba) as compared with rOmp25 and rOmp31 has been tested on the sera of 109 cattle with positive results to brucellosis according to classical serological tests. The rOmpBm-Ba had a higher antigenicity than the single ones, since it confirmed the presence of Brucella-antibody in all serum samples with positive results of i-ELISA/rOmp25 and/or i-ELISA/rOmp31, as well as additionally revealing 7.3% and 31.2% seropositive animals, respectively. A comparative study of the diagnostic value of i-ELISA/rOmpBm-Ba and conventional tests on blood sera of 24 cattle subjected to a bacteriological examination at slaughter showed a higher reliability of enzyme immunoassay. Further research is necessary to get a more objective evaluation of the diagnostic accuracy of Brucella rOmpBm-Ba by challenging laboratory animals.
8
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Application of selected medicinal plants such as chamomile and kalanchoe from in vitro culture for production of recombinant proteins

84%
Bandurska K., Krupa P.
BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
|
2015
|
tom 96
|
nr 1
9

Serodiagnosis of bovine lungworm infection using a recombinant protein as the capture antigen

84%
Hoglund J., Engstrom A., Mattsson J., Morrison D., Rydzik A.
Wiadomości Parazytologiczne. Suplement
|
2008
|
tom 54
10

The use of recombinant proteins in diagnostic parasitology: possibilities and challenges

84%
Mattsson J. G.
Wiadomości Parazytologiczne. Suplement
|
2008
|
tom 54
11
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Content available

Oral Lyme disease vaccine

84%
Urbanowicz A., Lewandowski D., Figlerowicz M.
BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
|
2014
|
tom 95
|
nr 4
12

Expression, purification, and bioactivity of a soluble recombinant ovine interferon-tau in Escherichia coli

84%
Yu H.-Y., Gao D.-M., Zhou W., Xia B.-B., He Z.-Y., Wu b., Jiang M.-Z., Wang M.-L., Zhao J.
Journal of Veterinary Research
|
2021
|
tom 65
|
nr 1
13

Recombinant C-reactive protein: a potential candidate for the treatment of cutaneous leishmaniasis of BALB/c mice caused by Leishmania major

84%
Zahedi S. N., Hejazi S. H., Boshtam M., Amini F., Fazeli H., Sarmadi M., Rahimi M., Khanahmad H.
Acta Parasitologica
|
2021
|
tom 66
|
nr 1
14
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Expression of the gene encoding blood coagulation factor VIII without domain B in E. coli bacterial expression system

84%
Mazurkiewicz-Pisarek A., Mazurkiewicz A., Mikiewicz D., Baran P., Ciach T.
BioTechnologia. Journal of Biotechnology Computational Biology and Bionanotechnology
|
2023
|
tom 104
|
nr 3
15
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Expression of gG, gD, gI and gE glycoprotein genes of Aujeszky's disease virus in baculovirus system and the studies of biological properties of recombinant proteins

84%
Bienkowska-Szewczyk K., Tyborowska J., Kochan G., Szymanski P., Szewczyk B.
Bulletin of the Veterinary Institute in Puławy
|
1998
|
tom 42
|
nr 1
16

Secretion of proteins by Escherichia coli and its application in production of recombinant proteins

84%
Fikus M. M., Rempola B.
Postępy Biochemii
|
1995
|
tom 41
|
nr 5
17

Chimeric protein ABRaA-VEGF121 is cytotoxic towards VEGFR-2-expressing PAE cells and inhibits B16-F10 melanoma growth

67%
Smagur A., Boyko M. M., Biront N. V., Cichon T., Szala S.
Acta Biochimica Polonica
|
2009
|
tom 56
|
nr 1
115-124
It has been known that VEGF121 isoform can serve as a carrier of therapeutic agents targeting tumor endothelial cells. We designed and constructed synthetic cDNA that encodes a chimeric protein comprising abrin-a (ABRaA) toxin A-chain and human VEGF121. Expression of the ABRaA-VEGF121 chimeric protein was carried out in E. coli strain BL21(DE3). ABRaA-VEGF121 preparations were isolated from inclusion bodies, solubilized and purified by affinity and ion-exchanged chromatography (Ni-agarose and Q-Sepharose). Finaly, bacterial endotoxin was removed from the recombinant protein. Under non-reducing conditions, the recombinant protein migrates in polyacrylamide gel as two bands (about 84 kDa homodimer and about 42 kDa monomer). ABRaA-VEGF121 is strongly cytotoxic towards PAE cells expressing VEGFR-2, as opposed to VEGFR-1 expressing or parental PAE cells. The latter are about 400 times less sensitive to the action of this fusion protein. The biological activity of the ABRaA domain forming part of the chimeric protein was assessed in vitro: ABRaA-VEGF121 inhibited protein biosynthesis in a cell-free translation system. Preincubation of ABRaA-VEGF121 with antibody neutralizing the biological activity of human VEGF abolished the cytotoxic effect of the chimeric protein in PAE/KDR cells. Experiments in vivo demonstrated that ABRaA-VEGF121 inhibits growth of B16-F10 murine melanoma tumors.
18

Cloning and expression of a new recombinant thrombolytic and antithrombotic agent - a staphylokinase variant

67%
Kowalski M., Brown G., Bieniasz M., Oszajca K., Chabielska E., Pietras T., Szemraj Z., Makandjou-Ola E., Bartkowiak J., Szemraj J.
Acta Biochimica Polonica
|
2009
|
tom 56
|
nr 1
41-53
 To develop a more potent antithrombin agent with thrombolytic and antiplatelet properties, a new staphylokinase (SAK) variant was constructed. The kringle 2 domain (K2) of tissue type-plasminogen activator (t-PA) containing a fibrin-specific binding site (i), the RGD sequence (Arg-Gly-Asp) for the prevention of platelet aggregation (ii) and the antithrombotic agent - hirulog (iii) was assembled to the C-terminal part of recombinant staphylokinase (r-SAK). cDNA for the hybrid protein SAK-RGD-K2-Hirul was cloned into Pichia pastoris pPIC9K yeast expression vector. The introduction of K2 t-PA, the RGD sequence and hirulog into the C-terminus of r-SAK did not alter the staphylokinase activity. We observed a higher clot lysis potency of SAK-RGD-K2-Hirul as evidenced by a faster and more profound lysis of 125I-labeled human fibrin clots. The potency of thrombin inhibition by the hirulog C-terminal part of the recombinant fusion protein was almost identical to that of r-Hir alone. These results suggest that the SAK-RGD-K2-Hirul construct can be a more potent and faster-acting thrombolytic agent with better antithrombin and antiplatelet properties compared to r-SAK and SAK-RGD-K2-Hir.
19

Characterisation of NcGRA7 and NcSAG4 proteins: immunolocalisation and their role in the host cell invasion by Neospora caninum tachyzoites

67%
Aguado-Martinez A., Alvarez-Garcia G., Schares G., Risco-Castillo V., Fernandez-Garcia A., Marugan-Hernandez V., Ortega-Mora L. M.
Acta Parasitologica
|
2010
|
tom 55
|
nr 4
304-312
Neospora caninum negatively impacts bovine reproductive performance around the world. Addressing this problem requires a greater understanding of the parasite’s molecular biology. In this study, monoclonal antibodies against recombinant proteins were successfully developed and employed to characterise two different proteins of N. caninum: the acute phase-associated NcGRA7 and the chronic phase-associated NcSAG4. Immunofluorescence with the anti-rNcGRA7 monoclonal antibody suggested that NcGRA7 trafficks from tachyzoite dense granules to the matrix of the parasitophorous vacuole and parasite’s surroundings. Furthermore, NcGRA7 is also expressed in the bradyzoite stage and localised on the matrix of bradyzoite-positive vacuoles. NcGRA7 appears to be partially involved in the tachyzoite-invasion mechanisms, as an anti-rNcGRA7 monoclonal antibody partially inhibited in vitro tachyzoite-invasion. A monoclonal antibody specific for NcSAG4 confirmed this protein’s bradyzoitespecific expression both by western blot and immunofluorescence. However, some bradyzoite-positive vacuoles only weakly expressed NcSAG4, if it was expressed at all. The specificity of the anti-rNcSAG4 monoclonal antibody was confirmed by the recognition of the NcSAG4 in the membrane surface of Nc-1SAG4c transgenic tachyzoites, which constitutively expresses NcSAG4. Blocking NcSAG4 of Nc-1SAG4c tachyzoites with the monoclonal antibody did not affect host cell invasion. However, its implication on the host cell adhesion or host immune evasion should not be discarded.
20

Transgenic technology as it applies to animal agriculture

67%
Kopchick J. J., Jura J., Mukerji P., Kelder B.
Biotechnologia
|
1996
|
nr 2
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