Dual priming oligonucleotide system for the multiplex detection of respiratory viruses and SNP genotyping of CYP2C19 gene

Successful PCR starts with proper priming between an oligonucleotide primer and the template DNA. However, the inevitable risk of mismatched priming cannot be avoided in the currently used primer system, even though considerable time and effort are devoted to primer design and optimization of reaction conditions. Here, we report a novel dual priming oligonucleotide (DPO) which contains two separate priming regions joined by a polydeoxyinosine linker. The linker assumes a bubble-like structure which itself is not involved in priming, but rather delineates the boundary between the two parts of the primer. This structure results in two primer segments with distinct annealing properties: a longer 5′-segment that initiates stable priming, and a short 3′-segment that determines target-specific extension. This DPO-based system is a fundamental tool for blocking extension of non-specifically primed templates, and thereby generates consistently high PCR specificity even under less than optimal PCR conditions. The strength and utility of the DPO system are demonstrated here using multiplex PCR and SNP genotyping PCR

Experimental Studies on Wave Interactions of Partially Perforated Wall under Obliquely Incident Waves

This study presents wave height distribution in terms of stem wave evolution phenomena on partially perforated wall structures through three-dimensional laboratory experiments. The plain and partially perforated walls were tested to understand their effects on the stem wave evolution under the monochromatic and random wave cases with the various wave conditions, incident angle (from 10 to 40 degrees), and configurations of front and side walls. The partially perforated wall reduced the relative wave heights more effectively compared to the plain wall structure. Partially perforated walls with side walls showed a better performance in terms of wave height reduction compared to the structure without the side wall. Moreover, the relative wave heights along the wall were relatively small when the relative chamber width is large, within the range of the chamber width in this study. The wave spectra showed a frequency dependency of the wave energy dissipation. In most cases, the existence of side wall is a more important factor than the porosity of the front wall in terms of the wave height reduction even if the partially perforated wall was still effective compared to the plain wall

Nucleotide sequence of the vmhA gene encoding hemolysin from Vibrio mimicus

AbstractThe structural gene (vmhA) of hemolysin from Vibrio mimicus (ATCC33653) was cloned and sequenced. The vmhA gene contains an open reading frame consisting of 2232 nucleotides which can code for a protein of 744 amino acids with a predicted molecular mass of 83 059. The similarity of amino acid sequence shows 81.6% identity with Vibrio cholerae El Tor hemolysin

Flavonol glycosides from the aerial parts of Aceriphyllum rossii and their antioxidant activities

The methanol extract obtained from the aerial parts ofAceriphyllum rossii (Saxifragaceae) was fractionated into ethyl acetate (EtOAc),n-BuOH and H2O layers through solvent fractionation. Repeated silica gel column chromatography of EtOAc andn-BuOH layers afforded six flavonol glycosides. They were identified as kaempferol 3-O-β-D-glucopyranoside (astragalin,1), quercetin 3-O-β-D-glucopyranoside (isoquercitrin,2), kaempferol 3-O-α-L-rhamnopyranosyl (1→6)-β-D-glucopyranoside (3), quercetin 3-O-α-L-rhamnopyranosyl (1→6)-β-D-glucopyrano-side (rutin,4), kaempferol 3-O-[α-L-rhamnopyranosyl (1→4)-α-L-rhamnopyranosyl (1→6)-β-D-glucopyranoside] (5) and quercetin 3-O-[α-L-rhamnopyranosyl (1→4)-α-L-rhamnopyranosyl (1→6)-β-D-glucopyranoside] (6) on the basis of several spectral data. The antioxidant activity of the six compounds was investigated using two free radicals such as the ABTS free radical and superoxide anion radical. Compound1 exhibited the highest antioxidant activity in the ABTS2,2-azinobis-(3-ethylbenzothiazoline-6-sulfonic acid) radical scavenging method. 100 mg/L of compound1 was equivalent to 72.1±1.4 mg/L of vitamin C, and those of compounds3 and5 were equivalent to 62.7±0.5 mg/L and 54.3±1.3 mg/L of vitamin C, respectively. And in the superoxide anion radical scavenging method, compound5 exhibited the highest activity with an IC50 value of 17.6 ± 0.3 μM. In addition, some physical and spectral data of the flavonoids were confirme

Sound waves delay tomato fruit ripening by negatively regulating ethylene biosynthesis and signaling genes

AbstractRegulation of tomato fruit ripening may help extend fruit shelf life and prevent losses due to spoilage. Here, tomato fruit were investigated whether sound treatment could delay their ripening. Harvested fruit were treated with low-frequency sound waves (1kHz) for 6h, and then monitored various characteristics of the fruit over 14-days at 23±1°C. Seven days after the treatment, 85% of the treated fruit were green, versus fewer than 50% of the non-treated fruit. Most of the tomato fruit had transitioned to the red ripening stage by 14 days after treatment. Ethylene production and respiration rate were lower in the sound-treated than non-treated tomatoes. Furthermore, changes in surface color and flesh firmness were delayed in the treated fruit. To investigate how sound wave treatment effects on fruit ripening, the expression of ethylene-related genes was analyzed by quantitative real-time RT-PCR analysis. The expression level of several ethylene biosynthetic (ACS2, ACS4, ACO1, E4 and E8) and ripening-regulated (RIN, TAGL1, HB-1, NOR, CNR) genes was influenced by sound wave treatment. These results indicated that sound wave treatment delays tomato fruit ripening by altering the expression of important genes in the ethylene biosynthesis and ethylene signaling pathways