Authors:Özgür Appak, Selçuk Türkel, Nuran Esen, and Ayşe Aydan Özkütük
to antibiotics. Additionally, the definition is necessary to enable the collection of epidemiological data. In this study, the polymerase chain reaction (PCR)-restrictionenzyme analysis (PRA) method and DNA sequence analysis method were used to
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.
Authors:Maryam Rahimpour Hesari, Ali Salehzadeh, and Reza Kazemi Darsanaki
Depending on 81 bp repeats, a strain analysis of PCR-RFLP products was performed with Hae III restrictionenzyme (Thermo Scientific, USA), where 10 μL of PCR product of coa gene was incubated with 6 U of the enzyme at 37 °C for 45 min
Authors:M. Hassani, M. Naghavi, M. Shariflou, and P. Sharp
A RFLP approach was used to investigate polymorphism of
-gliadin genes in
using a F
population from the cross of accessions AUS18913 and CPI110856. A set of 150 F
progenies was genotyped by acid polyacrylamide gel electrophoresis (A-PAGE) and only one recombinant line of
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
I was associated with the novel
-gliadin gene (T
) using a 1,200 bp DNA fragment of a
-gliadin gene as a gene-specific probe. The
-gliadin gene (T
) may be of interest for further studies relating storage proteins and wheat bread-making quality.
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.
Authors:Norma L. Calderón, Felipa Galindo-Mu?iz, Mireya Ortiz, B. Lomniczi, T. Fehervari, and L. H. Paasch
A Newcastle disease virus (NDV) isolated in Mexico and called Chimalhuacan strain was characterised by gene F restriction enzyme analysis and found to be a genotype II velogenic virus. Haematological evaluations and histological studies of bone marrow were conducted on chickens experimentally infected with the Chimalhuacan virus and on control chickens. Within 72 hours post infection (hpi), a 50% decrease in thrombocyte and monocyte counts and a complete cellular depletion in bone marrow islands were evident in the infected group. These findings suggest that the Chimalhuacan strain of NDV causes an early and severe damage of the haematopoietic cells including thrombocyte precursors, which might explain the marked thrombocytopenia detected in early stages of this disease.
Authors:Judit Pászti, Józsefné Király, and Miklós Füzi
Eighty isolates of Listeria monocytogenes cultured from human infections in Hungary between 2004 and 2012 were serotyped by the PCR technique of Doumith et al.  and characterised by pulsed-field gel electrophoresis (PFGE). Most of the isolates belonged to two serogroups: 53 isolates (66.3%) to serovar group 4b,4d,4e and 21 isolates (25.8%) to serogroup 1/2a,3a. Although many pulsotypes were identified a particular pulsotype proved highly excelling comprising of 31 isolates after digestion by both ApaI and AscI restriction enzymes. All strains from this pulsotype belonged to serovar group 4b,4d,4e. Interestingly 24% of isolates from invasive samples (cerebrospinal fluid, blood) belonged to two distinct pulsotypes in the less common serovar group 1/2a,3a. Several small clusters of cases caused by isolates with identical pulsotypes were identified.
Authors:Judit Bíró, Noémi Erdei, Ibolya Székely, and L. Stipkovits
Increasing use of Mycoplasma gallisepticum (MG) live vaccines has led to a need for the differentiation of MG strains. The MG strains MK-7, MS-16, S6, FS-9 and R strains and the MG live vaccine strain F were compared by random amplification of polymorphic DNA (RAPD) in this study. Using RAPD, different patterns were found among the MG strains. In addition to this, we examined the differentiating potential of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) primers targeted at the crmA, crmB, crmC, gapA, mgc2 and pvpA genes encoding cytadherence-related surface proteins. These proteins may take part in the pathogenesis of MG-induced disease. Differentiation of strain F is based on the identification of restriction enzyme sites in the PCR amplicons. Using HphI enzyme, crmC PCR amplicons produced different RFLP patterns. Digestion of amplicons of gapA-specific PCR with MboI enzyme also produced distinct patterns. Differences were observed among strains R and F by digestion of mgc2 PCR amplicons with HaeIII and VspI enzymes and digestion of pvpA PCR amplicons with AccI, PvuII and ScrFI endonucleases. This method can be used for the rapid differentiation of vaccine strain from wild strains. Differentiation of MG strains is a great advantage for diagnosticians or practitioners and it is useful for epidemiological studies.