• Journal of Microbiology and Biotechnology
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• v.28 no.9
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• pp.1547-1553
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• 2018

Hyaluronidases are a family of enzymes that catalyse the breakdown of hyaluronic acid, which is abundant in the extracellular matrix and cumulus oocyte complex. To investigate the activity of recombinant bovine sperm hyaluronidase 1 (SPAM1) and determine the effect of the Asn-X-Ser/Thr motif on its activity, the bovine SPAM1 open reading frame was cloned into the mammalian expression vector pCXN2 and then transfected to the HEK293 cell line. Expression of recombinant bovine hyaluronidase was estimated using a hyaluronidase activity assay with gel electrophoresis. Recombinant hyaluronidase could resolve highly polymeric hyaluronic acid and also caused dispersal of the cumulus cell layer. Comparative analysis with respect to enzyme activity was carried out for the glycosylated and deglycosylated bovine sperm hyaluronidase by N-glycosidase F treatment. Finally, mutagenesis analysis revealed that among the five potential N-linked glycosylation sites, only three contributed to significant inhibition of hyaluronic activity. Recombinant bovine SPAM1 has hyaluronan degradation and cumulus oocyte complex dispersion ability, and the N-linked oligosaccharides are important for enzyme activity, providing a foundation for the commercialization of hyaluronidase.

• Korean journal of applied entomology
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• v.53 no.4
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• pp.375-380
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• 2014

The sweetpotato whitefly (SPW), Bemisia tabaci Gennadius, is a major pest in tomato greenhouses on Jeju Island because they transmit viral diseases. To develop practical sampling methods for adult SPWs, yellow-color sticky traps were used in commercial tomato greenhouses throughout the western part of Jeju Island in 2011 and 2012. On the basis of the size and growing conditions in the tomato greenhouses, 20 to 30 traps were installed in each greenhouse for developing a sampling plan. Adult SPWs were more attracted to horizontal traps placed 60 cm above the ground than to vertical trap placed 10 cm above the plant canopy. The spatial patterns of the adult SPWs were evaluated using Taylor's power law (TPL) and Iwao's patchiness regression (IPR). The results showed that adult SPWs were aggregated in each surveyed greenhouse. In this study, TPL showed better performance because of the coefficient of determination ($r^2$). On the basis of the fixed-precision level sampling plan using TPL parameters, more traps were required for higher precision in lower SPW densities per trap. A sequential sampling stop line was constructed using TPL parameters. If the treatment threshold was greater than 10 maximum adult SPWs on a trap, the required traps numbered 15 at a fixed-precision level of 0.25. In estimating the mean density per trap, the proportion of traps with two or more adult SPWs was more efficient than whole counting: ${\ln}(m)=1.19+0.90{\ln}(-{\ln}(1-p_T))$. The results of this study could be used to prevent the dissemination of SPW as a viral disease vector by using accurate control decision in SPW management programs.