Wyniki wyszukiwania - Biblioteka Nauki

1

Cloning and characterization of the gene encoding ribosomal P0 phosphoprotein from Neurospora crassa.

100%

Krokowski D. , Tchórzewski M. , Grankowski N.

Acta Biochimica Polonica

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2002

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tom 49

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nr 1

11-18

EN

A gene for ribosomal protein P0 that belongs to the family of ribosomal P proteins was isolated from a Neurospora crassa cDNA library, using polyclonal antibodies against recombinant P0 protein from Saccharomyces cerevisiae. This is the first gene for ribosomal P0 protein to be cloned from filamentous fungi. The derived P0 protein sequence has a strong homology to other eukaryotic P0 proteins; yet, there is a notable alteration in the conservative C-terminal region, placing this protein among the unique sequences from protozoan parasites.

2

Animal cloning by nuclear transfer: state-of-the-art and future perspectives

100%

Dinnyes A. , Szmolenszky A.

Acta Biochimica Polonica

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2005

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tom 52

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nr 3

585-588

EN

Model organisms are essential to study the genetic basis of human diseases. Transgenic mammalian models, especially genetic knock-out mice have catalysed the progress in this area. To continue the advancement, further sophisticated and refined models are crucially needed to study the genetic basis and manifestations of numerous human diseases. Coinciding with the start of the new era of post-genomic research, new tools for establishment of transgenesis, such as nuclear transfer and gene targeting in somatic cells, have become available, offering a unique opportunity for the generation of transgenic animal models. The new technology provides important tools for comparative functional genomics to promote the interpretation and increase the practical value of the data generated in numerous mouse models. This paper discusses the state-of-the-art of the nuclear replacement technology and presents future perspectives.

3

An optimized recipe for cloning of the polymerase chain reaction-amplified DNA inserts into plasmid vectors.

100%

Topcu Z.

Acta Biochimica Polonica

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2000

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tom 47

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nr 3

841-846

EN

This study compares a number of parameters that are important in the ligation of the polymerase chain reaction-amplified DNA inserts into plasmid vectors and their efficient transformation to bacterial cells. The parameters covered were: T4 polynucleotide kinase treatment followed by either the large fragment of E. coli DNA polymerase or T4 DNA polymerase reactions, the amount of T4 DNA ligase, temperature and duration of ligation, molar ratio of insert to vector as well as the total DNA concentration. The results show that the T4 polynucleotide kinase-treated group without further enzymatic manipulation, at an insert to vector ratio of 3:1 gave the highest recombination efficiency when 10 μg/ml DNA and 20 units T4 DNA ligase were applied for ligation for 12 h at 4°C.

4

Molecular cloning and characterization of a peroxiredoxin from Phanerochaete chrysosporium

100%

Jiang Q. , Yan Y. H. , Hu G. K. , Zhang Y. Z.

Cellular and Molecular Biology Letters

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2005

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tom 10

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nr 4

5

An approach to cloning of Pi-b rice blast resistance gene

88%

Rybka K. , Miyamoto M. , Nakamura S. , Kawasaki S.

Acta Physiologiae Plantarum

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1997

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tom 19

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nr 4

EN

A contig of clones from BAC rice genomic library encompassing blast resistance gene Pi-b was constructed. On an average eight clones (8 ± 2.6) were picked up by each marker, which was expected basing on the BAC library size (Nakamura et al. 1997). The 2.4 cM distance between flanking RFLP markers G 1234 and RZ 213 (Miyamoto et al. 1996) was spanned with 4 steps of contig including 25 clones. The physical distance of 370 kb between flanking markers corresponds to a small ratio of physical and genetical distances (155 kb/cM) due to a probable structure of the gene locus near the telomeric end of the chromosome. Markers cosegregating with blast resistance against Magnoporthe grisea were localized in a 2 kb restriction fragment. A new border marker was found on the telomeric side of the Pi-b gene, less than 10 kb from cosegregating markers. No clear marker for the centromeric side of the gene was found but the position of Pi-b rice blast resistant gene was narrowed to within at least 50 kb, which is to our knowledge the most precised estimation of the position of this gene.

6

Molecular cloning and sequencing og the cDNA encoding plant 22 kDa nuclease

88%

Szmidzinski R. , Wilczynski G. , Szopa J.

Acta Biochimica Polonica

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1994

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tom 41

|

nr 2

7

Demethylation of the sex-determining region Y gene promoter and incidence of disorder of sex development in cloned dog males

75%

Hwang K. C. , Choi Y. K. , Jeong Y. I. , Park K. B. , Choi E. J. , Jeong Y. W. , Hossein M. S. , Hyun S. H. , Jeung E.-B. , Hwang W. S.

Journal of Physiology and Pharmacology

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2020

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tom 71

|

nr 3

8

Cloning, sequence, expression and characterization of human β-mannosidase

75%

Samra Z. Q. , Athar M. A.

Acta Biochimica Polonica

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2008

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tom 55

|

nr 3

479-490

EN

β-mannosidase (EC 3.2.1.25, MANB) dissects the non-reducing end of N-linked mannose moieties of glycoproteins in eukaryotic cells. The human β-mannosidase gene was amplified by RT-PCR, cloned and sequenced. The DNA sequence was compared with reported human β-mannosidase DNA sequence and sixteen nucleotide differences were found. The deduced amino-acid sequence showed that seven codons coded the same amino acids and nine codons coded different amino acids with reference to nucleotide substitution positions but did not affect recombinant MANB enzyme activity. No splice mutation was observed after comparison with reported MANB DNA sequences. A 75% homology of deduced amino-acid sequence was observed with mouse, goat and bovine β-mannosidase amino-acid sequences. The cloned β-mannosidase gene was subcloned into pET22b+ and pET28a+ expression vectors to transform the BL21-codon plus cells for expression of recombinant MAN22 and MAN28 enzymes, respectively. The optimized conditions for overexpression of recombinant β-mannosidase enzyme were induction with 1 mM IPTG for 12 h at 37°C. The expressed β-mannosidase enzyme was purified to homogeneity by a combination of DEAE-ion exchange and size exclusion chromatography. The molecular mass of MAN22 and MAN28 enzymes is 97 kDa by SDS/PAGE and is confirmed by western blot analysis. The recombinant enzymes are active at 37°C and at pH 5.0 and showed activity with p-nitrophenyl-β-d-mannopyranoside and not with p-nitrophenyl-α-d-mannopyranoside. The Km value of enzymes was 2.53 mM. The enzyme activity was inhibited by Zn2+, Co2+, Cu2+, Pb2+, Ag1+, iodoacetate, SDS, DMF, DMSO and ethanol. Fe3+, Ca2+ Mg2+, Mn2+, Triton X-100 and PMSF did not inhibit the enzyme activity. Northern blot analysis showed a transcript of about 3.7 kb in all cells and tissues studied. This is the first report on the expression and characterization of recombinant human MANB enzyme.

9

Cloning of two genes encoding Rab7 in Paramecium

75%

Surmacz L. , Wiejak J. , Wyroba E.

Acta Biochimica Polonica

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2006

|

tom 53

|

nr 1

149-156

EN

Rab7 is a small GTPase that plays a crucial role in the regulation of transport from early to late endosomes and lysosomes, phagosome maturation and in lysosomal biogenesis in mammalian cells. It contains conserved and unique sequence elements that mediate its function. Two Rab7 genes, Rab7a (703 bp) and Rab7b (707 bp) were identified in the unicellular eukaryote Paramecium by PCR amplification. They contain three short introns of different lengths (28-32 bp) and sequence located at identical positions in both genes. The presence of two Rab7 genes in the Paramecium genome was confirmed by Southern hybridization analysis performed with six different restriction enzymes. Expression of both genes was assessed by Northern blot and RT-PCR. Two transcripts of 1.8 and 2.2 kb were identified by hybridization analysis. The cloned complementary DNAs, both of 618 nucleotides in length, encode polypeptides of 206 amino acids that are 97.6% identical and differ in their C-termini. The predicted protein sequences of Rab7a and Rab7b contain all characteristic domains essential for Rab function: the effector domain (YRATVGADF) and four GTP-binding consensus sequences (GDSGVGKT, WDTAGQ, NKLD, SAK) as well as the prenylation motif (-CC) at the C-terminus indispensable for Rab binding to the membrane. Similarity searches revealed 81.6-82.1% homology of Paramecium Rab7 isoforms to human Rab7 and a lack of an insert typical for the Kinetoplastida - the species that appeared earlier in evolution. Paramecium is the first free-living lower eukaryote in which homologues of Rab7 have been identified that exhibit features similar to those of mammalian Rab7.

10

Bioinformatics analysis and expression of a novel protein ROP48 in Toxoplasma gondii

75%

Zhou J. , Wang L. , Zho A. , Lu G. , Li Q. , Wang Z. , Zhu M. , Zhou H. , Cong H. , He S.

Acta Parasitologica

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2016

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tom 61

|

nr 2

11

An organism arises from every nucleus

75%

Keklikoglu N.

Folia Histochemica et Cytobiologica

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2009

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tom 47

|

nr 2

179-183

12

Cloning and expression of apyrase gene from Ancylostoma caninum in Escherichia coli

75%

Qiao Y. , Pengsakul T.

Acta Parasitologica

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2015

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tom 60

|

nr 1

13

Cloning and characterization of M.LmoA118I, a novel DNA: m4C methyltransferase from the Listeria monocytogenes phage A118, a close homolog of M.NgoMXV

75%

Bujnicki J. M. , Radlinska M.

Acta Microbiologica Polonica

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2001

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tom 50

|

nr 2

14

Nuclear ribosomal DNA ITS paralogs as evidence of recennt interspecific hybridization in the genus Ophrys [Orchidaceae]

75%

Gulyas G. , Sramko G. , Molnar A. V. , Rudnoy S. , Illyes Z. , Balazs T. , Bratek Z.

Acta Biologica Cracoviensia. Series Botanica

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2005

|

tom 47

|

nr 2

EN

Ophrys holubyana Andrasovszky (Orchidaceae), distributed in the Carpatho-Pannonian region, is generally believed to be of hybrid origin. Although its hybrid origin is broadly accepted by many authors, no molecular evidence has been found to support the hypothesis. The nuclear ribosomal DNA internal transcribed spacer (nrDNA ITS) region was sequenced from Ophrys holubyana, and from the presumed progenitor taxa: Ophrys fuciflora (Cr.) Rchb. and Ophrys bicornis Sadler ex Nendtvich. Nearly all the known populations in the Carpatho-Pannonian region were sampled, and the first data on the nrDNA ITS sequence of O. holubyana and O. bicornis are presented. Aligning the ITS sequences revealed no differences among the ten samples. After cloning the amplified ITS regions, eight discrete ITS paralogs with regularly appearing nucleotide differences could be partitioned, differing in only 6 base pairs. Paralog sequences were detected not only in O. holubyana but also in some populations of the two parent species, suggesting that O. fuciflora and O. bicornis in the Carpatho-Pannonian region are also partially of hybrid origin themselves, or influenced by introgression. The study suggests that nrlTS regions can be generally useful in the study of Ophrys systematics and phylogeography and for analyzing the hybrid zones of related Ophrys species groups.

15

Molecular cloning and sequencing of the cDNA encoding plant nuclear matrix endonuclease

75%

Markiwicz E. , Rzepecki R. , Szopa J.

Acta Biochimica Polonica

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1994

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tom 41

|

nr 2

16

Reaction of potato clones with different type of resistance to potato virus M [PVM]

75%

Mietkiewska E.

Phytopathologia Polonica

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1994

|

tom 08

17

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

75%

Dabrowski S. , Kur J.

Acta Biochimica Polonica

|

1998

|

tom 45

|

nr 3

EN

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.

18

Cloning and expression of the genes coding for tube associated proteins of bacteriophage T4

75%

Nieradko J.

Acta Biochimica Polonica

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1995

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tom 42

|

nr 2

EN

Genes 29,48 and 54 of bacteriophage T4, coding for specific tube associated proteins, were cloned to the expression vector pT7-5. The molecular mass of the products of these genes was estimated to be 64,39 and 36 kDa, respectively. The examined genes are cotranscribed with genes 51,27 and 28 from the same DNA strand and a common late promoter sequence located downstream of gene 51.

19

Are there realistic opportunities to apply cloning and the other reproductive biotechnologies for safeguarding endangered species?

75%

Ptak G. , Loi P.

Animal Science Papers and Reports. Supplement

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2004

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tom 22

|

nr 1

20

Cloning of the Haemophilus influenzae Dam methyltransferase and analysis of its relationship to the Dam methyltransferase encoded by the HP1 phage

75%

Bujnicki J. M. , Radlinska M. , Zaleski P. , Piekarowicz A.

Acta Biochimica Polonica

|

2001

|

tom 48

|

nr 4

EN

In this paper we report cloning and experimental characterization of the DNA adenine methyltransferase (dam) gene from Haemophilus influenzae and comparison of its product with the Dam protein from the lysogenic phage of H. influenzae, HP1. Molecular modeling of M.HinDam and M.HP1Dam was carried out, providing a framework for a comparative analysis of these enzymes and their close homologs in the structural context. Both proteins share the common fold and essential cofactor-bind- ing and catalytic residues despite overall divergence. However, subtle but significant differences in the cofactor-binding pocket have been identified. Moreover, while M.HinDam seems to contact its target DNA sequence using a number of loops, most of them are missing from M.HP1Dam. Analysis of both MTases suggests that their catalytic activity was derived from a common ancestor, but similar sequence specificities arose by convergence.