Full-text resources of PSJD and other databases are now available in the new Library of Science.
Visit https://bibliotekanauki.pl
Preferences help
Polish version English version Language
enabled [disable] Abstract
Number of results

Results found: 14

Number of results on page
first rewind previous Page / 1 next fast forward last

Search results

help Sort By:

help Limit search:
first rewind previous Page / 1 next fast forward last
1
Content available remote

Identification and properties of the Deinococcus grandis and Deinococcus proteolyticus single-stranded DNA binding proteins (SSB)

100%
Filipkowski P., Kur J.
|
2007
|
vol. 54
|
issue 1
79-87
EN
To study the biochemical properties of SSB's from Deinococcus grandis (DgrSSB) and Deinococcus proteolyticus (DprSSB), we have cloned the ssb genes obtained by PCR and have developed Escherichia coli overexpression systems. The genes consist of an open reading frame of 891 (DgrSSB) and 876 (DprSSB) nucleotides encoding proteins of 296 and 291 amino acids with a calculated molecular mass of 32.29 and 31.33 kDa, respectively. The amino-acid sequence of DgrSSB exhibits 45%, 44% and 82% identity and the amino-acid sequence of DprSSB reveals 43%, 43% and 69% identity with Thermus aquaticus (TaqSSB), Thermus thermophilus (TthSSB) and Deinococcus radiodurans SSBs, respectively. We show that DgrSSB and DprSSB are similar to Thermus/Deinococcus SSBs in their biochemical properties. They are functional as homodimers, with each monomer encoding two single-stranded DNA binding domains (OB-folds). In fluorescence titrations with poly(dT), both proteins bind single-stranded DNA with a binding site size of about 33 nt per homodimer. In a complementation assay in E. coli, DgrSSB and DprSSB took over the in vivo function of EcoSSB. Thermostability with half-lives of about 1 min at 65-67.5°C make DgrSSB and DprSSB similar to the known SSB of Deinococcus radiodurans (DraSSB).
2
Content available remote

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

100%
Dąbrowski S., Kur J.
Acta Biochimica Polonica
|
1998
|
vol. 45
|
issue 3
661-667
3
Content available remote

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

100%
Dąbrowski S., Kur J.
Acta Biochimica Polonica
|
1998
|
vol. 45
|
issue 3
653-660
4
Content available remote

A freeze-thaw method for disintegration of Escherichia coli cells producing T7 lysozyme used in pBAD expression systems

81%
Wanarska M., Hildebrandt P., Kur J.
|
2007
|
vol. 54
|
issue 3
671-672
EN
The pLysN plasmid containing the T7 lysozyme gene under control of the lac promoter was constructed to facilitate cell disintegration after expression of recombinant proteins in arabinose-induced expression systems. The usefulness of this plasmid was tested in Escherichia coli TOP10 and E. coli LMG194 cells carrying pBADMHADgeSSB plasmid containing Deinococcus geothermalis SSB protein gene under control of the araBAD promoter. The results showed that low-level expression of T7 lysozyme did not interfere with the target SSB protein production, and that the freezing-thawing treatment was sufficient for disruption of the E. coli cells producing low amounts of T7 lysozyme.
5
Content available remote

Single-stranded DNA-binding proteins (SSBs) - sources and applications in molecular biology.

81%
Kur J., Olszewski M., Filipkowski P.
|
2005
|
vol. 52
|
issue 3
569-574
EN
Single-stranded DNA-binding proteins (SSBs) play essential roles in DNA replication, recombination, and repair in bacteria, archaea and eukarya. The SSBs share a common core ssDNA-binding domain with a conserved OB (oligonucleotide/oligosaccharide binding) fold. This ssDNA-binding domain was presumably present in the common ancestor to all three major branches of life. In recent years, there has been an increasing interest in SSBs because they are useful for molecular biology methods and for analytical purposes. In this review, we concentrate on recent advances in the discovery of new sources of SSBs as well as certain aspects of their applications in analytical sciences.
6
Content available remote

The chaperone-usher pathway of bacterial adhesin biogenesis - from molecular mechanism to strategies of anti-bacterial prevention and modern vaccine design*

81%
Piątek R., Zalewska B., Bury K., Kur J.
|
2005
|
vol. 52
|
issue 3
639-646
EN
The chaperone-usher system determines the biogenesis of surface-exposed adhesive structures responsible for virulence of many Gram-negative bacteria. Investigations of the last 20 years have resolved the mechanism of this pathway on a structural level for different species of pathogenic bacteria. The purpose of this review is to present the molecular mechanisms of the biogenesis of adhesive structures assembled via the chaperone-usher pathway. The obtained mechanistic data allow one to propose potential strategies of anti-bacterial action. Additionally, the specific properties of the polymeric adhesive structures (pili and fimbriae) of the chaperone-usher system allow their use as effective and safe recombinant vaccines carrying foreign epitopes in thousands of copies on bacterial cell surface.
7
Content available remote

Thermal profile with alternately raised and lowered annealing temperature improves the PCR amplification using highly degenerate primers

81%
Sachadyn P., Sobiewska G., Kur J.
Acta Biochimica Polonica
|
1998
|
vol. 45
|
issue 3
691-694
8
Publication available in full text mode
Content available

Principles and applications of Ligation Mediated PCR methods for DNA-based typing of microbial organisms

81%
Krawczyk B., Kur J., Stojowska-Swędrzyńska K., Śpibida M.
|
2016
|
vol. 63
|
issue 1
39-52
EN
A significant number of DNA-based techniques has been introduced into the field of microorganisms' characterization and taxonomy. These genomic fingerprinting methods were developed to detect DNA sequence polymorphisms by using general principles, such as restriction endonuclease analysis, molecular hybridization, and PCR amplification. In recent years, some alternative techniques based on ligation of oligonucleotide adapters before DNA amplification by PCR, known as Ligation-Mediated PCR methods (LM PCR), have been successfully applied for the typing of microorganisms below the species level. These molecular methods include: Amplified Fragment Length Polymorphism (AFLP), Amplification of DNA fragments Surrounding Rare Restriction Sites (ADSRRS), PCR Melting Profiles (PCR MP), Ligation Mediated PCR/Shifter (LM PCR/Shifter), Infrequent-Restriction-Site Amplification (IRS PCR), double digestion Ligation Mediated Suppression PCR (ddLMS PCR). These techniques are now applied more and more often because they involve less time, are comparably inexpensive, and require only standard lab equipment. Here, we present a general review of this group of methods showing their possibilities and limitations. We also identify questions and propose solutions which may be helpful in choosing a particular LM PCR method for the achievement of the required goal.
9
Content available remote

Thermostable Pyrococcus woesei β-D-galactosidase - high level expression, purification and biochemical properties

81%
Wanarska M., Kur J., Pladzyk R., Turkiewicz M.
|
2005
|
vol. 52
|
issue 4
781-787
EN
The gene encoding β-D-galactosidase from Pyrococcus woesei was PCR amplified, cloned, expressed in Escherichia coli under the control of an inducible T7 promoter, purified and characterized. The expression system was developed by the construction of recombinant plasmid, based on the high copy number pUET1 vector, giving four times more efficient expression of P. woesei β-D-galactosidase (20 mg of enzyme from 1 liter of culture) than that obtained from a previously constructed one. The recombinant enzymes were purified in a two-step procedure: double heat-denaturation of E. coli cell proteins and affinity chromatography on p-aminobenzyl 1-thio-β-D-galactopyranoside-agarose. To achieve efficient purification of P. woesei β-D-galactosidase by immobilized metal-ion affinity chromatography (IMAC), a His-tag was placed either at the N- or the C-terminal of the coding sequence. The obtained fusion proteins revealed the same specific activity of approximately 5400 U/mg, which was 10 times lower than the wild-type β-D-galactosidase (51100 U/mg). The activity of P. woesei β-D-galactosidase was enhanced by thiol compounds, Mg2+ ions and D-galactose, and was inhibited by heavy metal ions and D-glucose, while Ca2+ ions had no effect.
10
Content available remote

Binding of Escherichia coli integration host factor (IHF) to the origin segment of p15A plasmid

80%
Hiszczyńska-Sawicka E., Kur J.
Acta Biochimica Polonica
|
1995
|
vol. 42
|
issue 1
103-108
11
Publication available in full text mode
Content available

Cloning, expression, and biochemical characterization of a cold-active GDSL-esterase of a Pseudomonas sp. S9 isolated from Spitsbergen island soil

62%
Wicka M., Wanarska M., Krajewska E., Pawlak-Szukalska A., Kur J., Cieśliński H.
|
2016
|
vol. 63
|
issue 1
117-125
EN
An estS9 gene, encoding an esterase of the psychrotolerant bacterium Pseudomonas sp. S9 was cloned and sequenced. The deduced sequence revealed a protein of 636 amino acid residues with a molecular mass of 69 kDa. Further amino acid sequence analysis revealed that the EstS9 enzyme contained a G-D-S-L motif centered at a catalytic serine, an N-terminal catalytic domain and a C-terminal autotransporter domain. Two recombinant E. coli strains for production of EstS9N (a two domain enzyme) and EstS9Δ (a one domain enzyme) proteins were constructed, respectively. Both recombinant proteins were successfully produced as inclusion bodies and then purified under denaturing conditions. However, because of the low enzymatic activity of the refolded EstS9Δ protein, only the EstS9N protein was further characterized. The purified and refolded EstS9N protein was active towards short-chain p-nitrophenyl esters (C2-C8), with optimal activity for the butyrate (C4) ester. With p-nitrophenyl butyrate as the substrate, the enzyme displayed optimal activity at 35°C and pH 9.0. Additionally, the EstS9N esterase retained ~90% of its activity from 25-40°C and ~40% of its activity at 10°C. Moreover, analysis of its kinetic parameters (Km, kcat, kcat/Km) toward p-nitrophenyl butyrate determined at 15°C and 25°C confirmed that the EstS9 enzyme is cold-adapted. To the best of our knowledge, EstS9 is the third characterized cold-active GDSL-esterase and the first one confirmed to contain an autotransporter domain characteristic for enzymes secreted by the type V secretion system.
12
Content available remote

Construction of a DNA-polymerase I overproducing plasmid and isolation of the enzyme

62%
Bielawski K., Skowron P., Kur J., Konopa G., Podhajska A., Taylor K.
|
|
issue 1
29-34
13
Content available remote

Protection of cattle against bovine leukemia virus (BLV) infection could be attained by DNA vaccination

62%
Brillowska A., Dąbrowski S., Rułka J., Kubiś P., Buzała E., Kur J.
Acta Biochimica Polonica
|
1999
|
vol. 46
|
issue 4
971-976
EN
The bovine leukemia virus (BLV) envelope gene encoding extracellular glycoprotein gp51 and transmembrane glycoprotein gp30 was cloned into a vehicle expression vector under the human cytomegalovirus (CMV) intermediate early promoter. The intramuscular injection of this plasmid vector generated a cellular immune response. Seven out of ten cows vaccinated with the DNA construct resisted a drastic challenge (500 BLV-infected lymphocytes as an infectious dose).
14
Content available remote

Purification of IHF-like protein from gram-negative bacteria in one chromatographic step

60%
Krawczyk B., Kur J.
Acta Biochimica Polonica
|
1996
|
vol. 43
|
issue 2
379-382
first rewind previous Page / 1 next fast forward last
JavaScript is turned off in your web browser. Turn it on to take full advantage of this site, then refresh the page.