• Applied Biological Chemistry
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• v.37 no.1
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• pp.56-63
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• 1994

Oligonucleotides for a conserved region of the coat protein gene of cucumber mosaic virus (CMV) and a hammerhead structure ribozyme against CMV RNA were synthesized using a DNA synthesizer. Both strands of oligonucleotides were annealed and restricted with BamHI/SacI, then cloned into a plasmid pBS SK (+). The cloned CMV substrate and ribozyme were sequenced to verify correct constructions. In vitro transcriptions were carried out by using T7 RNA polymerase with BssHII or SspI digests of $1\;{\mu}g$ of substrate and ribozyme clones. The size of substrate RNA was 176 nucleotides (nt) containing 50 nt of CMV RNA sequence, 6 nt of XbaI restriction site and 120 nt of vector-derived sequence in the case of BssHII digest. The size of ribozyme RNA was 164 nt containing 40 nt of ribozyme RNA sequence and same sequences of substrate. Substrate RNA was efficiently cleaved into two fragments (96 nt and 80 nt) by ribozyme RNA. This endonucleolytic cleavage occurred more efficiently at $55^{\circ}C$ than $37^{\circ}C$. SspI digest-derived substrate RNA (2234 nt) was also cleaved into two fragments by the same ribozyme. SspI digest-derived ribozyme RNA (2222 nt) cleaved the above substrate to two fragments. In vitro-tested ribozyme construct is being cloned into a plant transformation vector to develop virus-resistant plants.

• Journal of the Korea Organic Resources Recycling Association
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• v.23 no.1
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• pp.70-81
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• 2015

Helicases are known to be a proteins that use the chemical energy of NTP binding and hydrolyze to separate the complementary strands of double-stranded nucleic acids to single-stranded nucleic acids. They participate in various cellular metabolism in many organisms. DEAD-box proteins are ATP-dependent RNA helicase that participate in all biochemical steps involving RNA. DEAD-box3 (DDX3) gene is belonging to the DEAD-box family and plays an important role in germ cell development in many organisms including not only vertebrate, but also invertebrate during asexual and sexual reproduction and participates in stem cell differentiation during regeneration. In this study, in order to identify and characterize DDX3 gene in the earthworm, Perionyx excavatus having a powerful regeneration capacity, total RNA was isolated from adult head containing clitellum. Full length of DDX3 gene from P. excavatus, Pe-DDX3, was identified by RT-PCR using the total RNA from head as a template. Pe-DDX3 encoded a putative protein of 607 amino acids and it also has the nine conserved motifs of DEAD-box family, which is characteristic of DEAD-box protein family. It was confirmed that Pe-DDX3 has the nine conserved motifs by the comparison of entire amino acids sequence of Pe-DDX3 with other species of different taxa. Phylogenetic analysis revealed that Pe-DDX3 belongs to a DDX3 (PL10) subgroup of DEAD-box protein family. And it displayed a high homology with PL10a, b from P. dumerilii.

• Asian Pacific Journal of Cancer Prevention
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• v.17 no.7
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• pp.3309-3315
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• 2016

Helicobacter pylori as the second most common cause of gastric cancer in the world infects approximately half of the developed countries population and 80% of the population living in developing countries. Integrons as genetic reservoirs play major roles in dissemination of antimicrobial resistance genes. To the best of our knowledge, this is the first study to report carriage of class 1 and 2 integrons and associated gene cassettes in H. pylori isolates from Iran. This cross-sectional study was conducted in Tehran among 110 patients with H. pylori infection. Antimicrobial susceptibility testing (AST) for H. pylori strains were assessed by the micro broth dilution method. Class 1 and 2 integrons were detected using PCR. In order to determine gene cassettes, amplified fragments were subjected to DNA sequencing of both amplicon strands. The prevalence of resistance to clarithromycin, metronidazole, clarithromycin, tetracycline, amoxicillin, rifampin, and levofloxacin were 68.2% (n=75), 25.5% (n=28), 24.5% (n=27), 19.1% (n=21), 18.2% (n=20) and 16.4% (n=18), respectively. Frequency of multidrug resistance among H. pylori isolates was 12.7%. Class 2 integron was detected in 50 (45.5%) and class 1 integron in 10 (9.1%) H. pylori isolates. The most predominant gene cassette arrays in class 2 integron-bearing H. pylori were included sat-era-aadA1, dfrA1-sat2-aadA1, blaoxa2 and, aadB whereas common gene cassette arrays in class 1 integron were aadB-aadA1-cmlA6, aacA4, blaoxa2, and catB3. The high frequency of class 2 integron and multidrug resistance in the present study should be considered as a warning for clinicians that continuous surveillance is necessary to prevent the further spread of resistant isolates.

• Journal of Korean Society of Forest Science
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• v.47 no.1
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• pp.1-15
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• 1980

The occurrence of witches' broom of Rhus javanica was first noticed in Korea by the author in 1979. Subsequently, studies were made on the symptomatology, etiology, and transmission of the disease, as well as the effect of some antibiotics on the disease development. The results of these studies are summarized as follows: 1. Symptoms of the infected plant were characterized by dwarfing of the tree accompanied by yellowing and brooming of the foliage. 2. Electron microscopy of witches' broom diseased Rhus javanica plant revealed the occurrence of numerous mycoplasma-like organisms (MLO's) in the phloem tissue cells (sieve tube elements and phloem parenchyma cells) of the rachis and midribs of infected leaves. 3. The MLO's were bounded by a single unit membrane and contained ribosome-like granules and strands presumed to be DNA. It also appears that the MLO multiply possibly by budding as well as binary and plurinary fission. 4. In the midrib of healthy leaves, vascular bundles were collaterally discontinuous. In the diseased leaves, however, xylems were connected to each other and phloem cells showed an atrophy. Granules, which were prominent in the normal abaxial epidermis, were not observed in the peidermis of diseased leaves. 5. Electron microscopy revealed crystals or osmopholic granules in the phloem parenchyma cells, and that normal stacks of grana were not developed in the chloroplasts of infected levels. 6. The disease was experimentally transmitted by grafting. Budding was more effective than crown grafting for transmitting the disease. The disease has been transmitted by grafting even when complete union of stocks and scions has not taken place. The disease agent was not transmitted by sap inoculation. Insect transmission has not been confirmed. 7. Dipping the roots of infected plants into the 500 ppm and 1,000 solutions of either tetracycline HCI or oxytetracycline, HCI was more effective on temporary remision of the symptoms than spraying the 100 ppm and 200 ppm solutions of the same antibiotics. A greater effect was achieved through dipping into 1,000 ppm than into 500 ppm.