Search Results

You are looking at 1 - 8 of 8 items for :

  • "restriction enzymes" x
  • Biology and Life Sciences x
  • All content x
Clear All
Acta Biologica Hungarica
Authors: Edit Nádasi, P. Gyűrűs, Márta Czakó, Judit Bene, Sz. Kosztolányi, Sz. Fazekas, P. Dömösi, and B. Melegh

Hungarians are unique among the other European populations because according to history, the ancient Magyars had come from the eastern side of the Ural Mountains and settled down in the Carpathian basin in the 9th century AD. Since variations in the human mitochondrial genome (mtDNA) are routinely used to infer the histories of different populations, we examined the distribution of restriction fragment length polymorphism (RFLP) sites of the mtDNA in apparently healthy, unrelated Hungarian subjects in order to collect data on the genetic origin of the Hungarian population. Among the 55 samples analyzed, the large majority belonged to haplogroups common in other European populations, however, three samples fulfilled the requirements of haplogroup M. Since haplogroup M is classified as a haplogroup characteristic mainly for Asian populations, the presence of haplogroup M found in approximately 5% of the total suggests that an Asian matrilineal ancestry, even if in a small incidence, can be detected among modern Hungarians.

Restricted access

Genetic/genomic polymorphism, i.e. variations in DNA sequences are ideally assayed by direct nucleotide sequencing of a gene region or other homologous segment of the genome. An easier and cheaper approach, however, if the variants are analyzed by hybridization technology using restriction fragment length polymorphisms (RFLPs) or by detection of the number of tandem repeats (VNTR) of small DNA segments, the “minisatellites”. In this study we describe results of the DNA analysis of repetitive sequences of human 6th chromosome by the application of a chemiluminescent labeled probes. The allele frequency distribution of polymorphic DNA sequences has been determined in unrelated individuals. The isolated genomic DNA was cut with Pst I restriction enzyme, size fractionated on agarose gel and hybridized with a chemiluminescent labeled D6 S132 probe. At this locus the Pst I cleaved DNA fragments are ranging from 1841 to 6098 base pairs (bp). Specific genetic pattern was characterized by more frequent fragments (3313 and 3884 bp), and the rarely occurring ones (clustered between 1841-2595 and 5227-6098 bp). Our study provides a further possibility for characterization of individual genomic patterns.

Restricted access

A RFLP approach was used to investigate polymorphism of ω -gliadin genes in Ae. tauschii using a F 2 population from the cross of accessions AUS18913 and CPI110856. A set of 150 F 2 progenies was genotyped by acid polyacrylamide gel electrophoresis (A-PAGE) and only one recombinant line of Gli-D t 1/Gli-D t T1 was observed. Twelve restriction enzymes were initially tested on genomic DNA of the two parents of which four restriction enzymes revealed polymorphism. Of these four, only Dra I was associated with the novel ω -gliadin gene (T 1 ) using a 1,200 bp DNA fragment of a ω -gliadin gene as a gene-specific probe. The ω -gliadin gene (T 1 ) may be of interest for further studies relating storage proteins and wheat bread-making quality.

Restricted access

The tuf gene of “Candidatus Phytoplasma mali”, the causal agent of apple proliferation was PCR cloned in an expression vector and expressed in Escherichia coli. First, phytoplasma DNA extracted from periwinkle was amplified using primers designed on the basis of the tuf gene and the PCR product was cloned into pGEM-T (Promega). In the next step specific primers were constructed containing some plasmid sequences and restriction enzyme sites. With this primers the sequence in pGEM-T was amplified, the product was digested with restriction enzymes, and inserted into the pQE40 expression vector (Qiagen). In this plasmid the tuf gene was fused to the 6xHIS tag, and DHFR. The production of 6xHIS-DHFR-Tu fusion protein protein was induced with IPTG and expressed in E. coli M15. The new fusion protein was found in the insoluble fraction of the bacterium. The identity of the protein was verified with polyacrylamid gel-electrophoresis and Western blot analysis using antiserum raised against the 6xHIStag of the fusion protein.

Restricted access

In PI466495, a powdery mildew resistance source of wild barley ( Hordeum vulgare ssp. spontaneum ), one gene conferring powdery mildew resistance was identified in the Mla locus. In this paper, the RGH1a gene sequence was used as source for the development of a cleaved amplified polymorphic sequence (CAPS) marker. Co-segregation between this marker and powdery mildew resistance was analysed by specific DNA fragments associated with each allele of the gene using 286 F 2 plants derived from a cross between winter barley ( H. vulgare L.) variety ‘Tiffany’ and PI466495. For the co-dominant marker RGH1aI1a , three fragments, 370 bp, 82 bp and 59 bp in size, were amplified from F 2 plants exhibiting resistance reaction types 0 and 0–1 to powdery mildew; whereas two fragments, 429 bp and 82 bp in size, were amplified in susceptible plants. Simple procedures based on polymerase chain reaction and restriction enzyme digestion allowed for identifying the plants susceptible to powdery mildew ( Blumeria graminis f. sp. hordei ) and plants homozygous or heterozygous for the resistance allele. The RGH1aI1a marker was positioned 0.85 cM to the resistance gene and the efficiency of marker-assisted selection (MAS), evaluated as the probability of crossing-over between the marker and the targeted gene, was 99%. The CAPS marker RGH1aI1a is a valuable candidate for MAS and gene transfer into barley varieties susceptible to powdery mildew.

Restricted access

41 Messelson, M., Yuan, R. 1968. DNA restriction enzyme from E. coli . Nature 217 :1110–1114. Yuan R. DNA

Restricted access
Biologia Futura
Authors: Muhammad Saad, Helen Mary, Umar Amjid, Ghulam Shabir, Kashif Aslam, Shahid Masood Shah, and Abdul Rehman Khan

restriction enzyme and the other subsample was digested with Hpa II by incubation at 37 °C overnight and enzymes inactivation at 80 °C for 20 min. The restricted samples were then ligated with Eco RI and Hpa II linkers (Table  1 ) using T4 DNA ligase enzyme

Restricted access

use of the Prime Script RT reagent Kit with gDNA Eraser (Takara, Japan) with following the operating instructions. The specific primers with cleavage sites (PstI and EcoRI) of restriction enzyme introduced, RcFATA-F (5′-AAAACTGCAGATGTTAAAAGTACCTTGTTG-3

Restricted access