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Vikky RAJULAPATI,Seunghyun HAN,Yong Hwan KIM 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.4
Lignin is the amplest sustainable aromatic polymer on earth. Lignin plays a key role in plants, provide the protect matrix to cellulose microfibrils of cell wall and helps to transport system in plants [1]. Its conversion to the value-added products from biomass waste research is increasing nowadays. Lignin degradation occurs oxidation by lignin peroxidases (EC 1.11.1.14, LiPs) in presence of H2O2. LiPs are the first class of enzymes to degrade lignin. The lignin peroxidases have various applications such as biorefinery, textile, energy, bioremediation, cosmetology and dermatology industries. LiPs are majorly secreted by white rot basidiomycete Phanerochaete chrysosporium, (PcLiPs). PcLiP isoenzymes have huge demand for industrial applications [2]. The production of PcLiPs have been showed low expression, yield of protein, incorrect fold and forming inclusion bodies in E. coli expression system. The recovery of LiPs using the unfolding and refolding by urea, which leads the expensive, time consumption process and ultimately the low production yield [3]. To overcome these, the present research approach, the PcLiP isoenzymes expression and production in eukaryotic system using different forms of P<sup>PICZ</sup> and P<sup>PICZα</sup> vectors through various Pichia pastoris strains. P. pastoris is widely used for production of heterologous proteins, due to posttranslational modifications, proper folding and high cell density [4]. P. pastoris is a suitable system for express and production of high-level secretion, PcLiP isoenzymes for industry applications.
Vikky RAJULAPATI,Seung-hyun HAN,Yong Hwan KIM 한국생물공학회 2021 한국생물공학회 학술대회 Vol.2021 No.10
Pichia pastoris is methylotrophic yeast and a commonly used eukaryotic host system for recombinant protein expression. Due to its posttranslational modifications, proper folding and high cell density. Lignin peroxidases (EC 1.11.1.14, LiPs), well known enzymes responsible for lignin degradation and a variety of aromatic pollutants such as phenolic and non-phenolic substrates by oxidize process. The lignin-degrading enzymes naturally produced by white-rot fungus, Phanerochaete chrysosporium. LiPs are the major class of enzymes to degrade lignin [2] and play vital role in cosmetics as skin whitening agents (melanin). The present study, gene encoding LiPs (1125 bp) from Phanerochaete chrysosporium codon optimised and cloned into methanol oxidise promoter vector PPICZα. The cloned vector is confirmed by sequence analysis and further linearized by Sac I restriction enzyme. The linearized vector transforms into Pichia pastoris strain X-33 genome by electroporation at 1500 V, 50 ㎌ and 200 Ω using a Gene Pulser II apparatus in 0.2 ㎝ cuvette and spread on 100 ㎍/mL zeocin YPDS plate incubated for 3-4 days at 30℃. The colonies further grew on higher zeocin concentrations (500, 1000, 1500 & 2000 ㎍/mL) for selection of multi copy recombinant. The selection of phenotypic screening was done using minimal glucose and methanol medium plates. Integration of LiP gene was confirmed by colony PCR with AOX1 primers. Lips gene expression of mRNA level studied by RTPCR using gene specific primers and standard genes. ACT gene used as normalization and endogenous expression. The expression level compared with native vector recombinant, wild type stain and without sample used as negative control. mRNA level of LiPs gene copy number increased as compared to control. LiPs expression will be studied on glycerol and methanol medium.
Chiu, Joanne,Tang, Vikki,Leung, Roland,Wong, Hilda,Chu, Kin Wah,Poon, Jensen,Epstein, Richard J.,Yau, Thomas Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.11
Background: Although FOLFOX (infusional fluorouracil/leucovorin plus oxaliplatin) is established as a standard chemotherapeutic regimen, the long term efficacy of adjuvant XELOX (oral capecitabine plus intravenous oxaliplatin) in Asian colorectal cancer (CRC) patients remains anecdotal. Moreover, uncertainties persist as to whether pharmacogenetic differences in Asian populations preclude equally tolerable and effective administration of these drugs. Method: One hundred consecutive patients with resected colorectal cancer received adjuvant XELOX (oxaliplatin 130 $mg/m^2$ on day 1 plus capecitabine 900 $mg/m^2$ twice daily on day 1 to 14 every 3 weeks for 8 cycles) at Queen Mary Hospital, Hong Kong. Endpoints monitored during follow-up were disease-free survival (DFS) and disease recurrence, overall survival (OS) and adverse events (AEs). Results: The median patient age was 56 years, 56% were diagnosed with rectal cancer and 44% with colonic cancer. After a median follow-up of 4.3 years (95% confidence interval, 3.2-4.7), 24 recurrences were confirmed including 13 patients who died due to progressive disease. Four-year DFS was 81% in colon cancer patients and 67% in rectal cancer patients (p=0.06 by log-rank test). For the cohort as a whole, OS was 90% at 3 years and 84% at 5 years. Treatment-related AEs led to early withdrawal in four patients. The commonest non-hematological AEs were neuropathy (91%), hand-foot syndrome (49%) and diarrhea (46%), while the commonest grade 3/4 AEs were neutropenia (11%) and diarrhea (10%). Conclusion: These results confirm the favourable long term survival benefit with good tolerability in using adjuvant XELOX in treating East Asian colorectal cancer patients.
Competition of sorghum cultivars and densities with Japanese millet (Echinochloa esculenta)
Wu, Hanwen,Walker, Steven R.,Osten, Vikki A.,Robinson, Geoff The Korean Society of Weed Science and The Turfgra 2010 Weed Biology and Management Vol.10 No.3
Field studies were conducted at two locations in southern Queensland, Australia during the 2003-2004 and 2004-2005 growing seasons to determine the differential competitiveness of sorghum (Sorghum bicolor L. Moench) cultivars and crop densities against weeds and the sorghum yield loss due to weeds. Weed competition was investigated by growing sorghum in the presence or absence of a model grass weed, Japanese millet (Echinochloa esculenta). The correlation analyses showed that the early growth traits (height, shoot biomass, and daily growth rate of the shoot biomass) of sorghum adversely affected the height, biomass, and seed production of millet, as measured at maturity. "MR Goldrush" and "Bonus MR" were the most competitive cultivars, resulting in reduced weed biomass, weed density, and weed seed production. The density of sorghum also had a significant effect on the crop's ability to compete with millet. When compared to the density of 4.5 plants per $m^2$, sorghum that was planted at 7.5 plants per $m^2$ suppressed the density, biomass, and seed production of millet by 22%, 27% and 38%, respectively. Millet caused a significant yield loss in comparison with the weed-free plots. The combined weed-suppressive effects of the competitive cultivars, such as MR Goldrush, and high crop densities minimized the yield losses from the weeds. These results indicate that sorghum competition against grass weeds can be improved by choosing competitive cultivars and by using a high crop density of >7.5 plants per $m^2$. These non-chemical options should be included in an integrated weed management program for better weed management, particularly where the control options are limited by the evolution of herbicide resistance.
Wei, Celine,Wilson, Carlene,Knott, Vikki Asian Pacific Journal of Cancer Prevention 2013 Asian Pacific journal of cancer prevention Vol.14 No.5
Background: The study investigated the influence of culturally-based health beliefs on engagement in healthy lifestyle behaviour. Specifically, the study compared levels of engagement between Western and Chinese youth in Australia and assessed the extent to which culture-specific attributions about the causes of illness, and health beliefs, predict engagement in healthy lifestyle behaviour. Materials and Methods: Ninety-four Western and 95 Chinese (N=189; Mean Age=20.8 years, SD=3 years) young adults completed an online questionnaire. Predictor variables were cultural health beliefs measured by the Chinese Cultural Views on Health and Illness scale (CCVH, Liang et al., 2008), and illness attributions beliefs measured by the Cause of Illness Questionnaire (CIQ, Armstrong and Swartzman, 1999). Outcomes variables were levels of engagement in healthy lifestyle behaviour. Results: Results indicated that Chinese participants have a significantly lower exercising rate and healthy dietary habits compared to the Western sample. Moreover, Chinese participants were found to believe more strongly than Westerners that cancer was associated with factors measured by the Traditional-Chinese-Model (TCM). Finally, the observed relationship between cultural health beliefs and physical inactivity was mediated by attributions of illness, in particular to the supernatural subscale, with the Sobel Test showing a significant mediation (z=-2.63, p=0.004). Conclusions: Mainstream approaches to encourage healthy lifestyles are unlikely to be effective when educating Chinese youth. Instead, health promotion programs should attempt to address the illness attribution beliefs and educate Chinese youth about the role of diet and exercise in prevention of diseases such as cancer.