• 한국동물생명공학회지
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• 제34권4호
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• pp.280-291
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• 2019

Potential utility of 14 candidate housekeeping genes as normalization reference for RT-qPCR analysis with developmental samples (fertilized eggs to late veliger larvae) in Pacific abalone Haliotis discus hannai was evaluated using four different statistical algorithms (geNorm, NormFinder, BestKeeper and comparative ΔCT method). Different algorithms identified different genes as the best candidates, and geometric mean-based final ranking from the most to the least stable expression was as follow: RPL5, RPL4, RPS18, RPL8, RPL7, UBE2, RPL7A, GAPDH, RPL36, PPIB, EF1A, ACTB and B-TU. The findings were further validated via relative quantification of metallothionein (MT) transcripts using the stable and unstable reference genes, and expression levels of MT were greatly influenced according to the choice of reference genes. In overall, our data suggest that RPL5 and RPS18, either singly or in combination, are appropriate for normalizing gene expression in developmental samples of this abalone species, whereas ACTB, B-TU and EF1A are less stable and not recommended. In addition, our findings propose that standard deviations in geometric ranking as well as geometric mean itself should also be taken into account for the final selection of reference gene(s). This study could be a useful basis to facilitate the generation of accurate and reliable RT-qPCR data with developmental samples in this abalone species.

• Molecules and Cells
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• 제25권2호
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• pp.258-264
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• 2008

Deschampsia antarctica is the only monocot that thrives in the tough conditions of the Antarctic region. It is an invaluable resource for the identification of genes associated with tolerance to various environmental pressures. In order to identify genes that are differentially regulated between greenhouse-grown and Antarctic field-grown plants, we initiated a detailed gene expression analysis. Antarctic plants were collected and greenhouse plants served as controls. Two different cDNA libraries were constructed with these plants. A total of 2,112 cDNA clones was sequenced and grouped into 1,199 unigene clusters consisting of 243 consensus and 956 singleton sequences. Using similarity searches against several public databases, we constructed a functional classification of the ESTs into categories such as genes related to responses to stimuli, as well as photosynthesis and metabolism. Real-time PCR analysis of various stress responsive genes revealed different patterns of regulation in the different environments, suggesting that these genes are involved in responses to specific environmental factors.

• 한국축산식품학회지
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• 제29권2호
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• pp.157-163
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• 2009

This study was conducted for the isolation, purification, and characterization of a protease from Korean pear, to see its proteolytic activity on chicken actomyosin and to find the optimum pH and temperature of activity on chicken actomyosin. The protease was isolated from crude extract of Korean pear by ammonium sulfate precipitation. Further purification was done by DEAE-Sepharose ion-exchange chromatography, Mono-Q and Mini-Q column chromatography. The purified enzyme gave a single protein band on SDS polyacrylamide gel electrophoresis and the molecular weight was found to be 38 kDa. The specific activity of purified enzyme was 34,907 unit/mg with 25 fold purification and the yield was 2%. The purified enzyme incubated with chicken actomyosin showed high activity. The optimum pH and temperature for enzyme activity on chicken actomyosin were 6.5 and $70^{\circ}C$, respectively. A protease was purified from Korean pear for the first time and characterized. It was found to be promising for meat tenderization.

• Asian-Australasian Journal of Animal Sciences
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• 제26권12호
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• pp.1665-1671
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• 2013

Real-time quantitative PCR (qRT-PCR) is one of the important methods for investigating the changes in mRNA expression levels in cells and tissues. Selection of the proper reference genes is very important when calibrating the results of real-time quantitative PCR. Studies on the selection of reference genes in goat tissues are limited, despite the economic importance of their meat and dairy products. We used real-time quantitative PCR to detect the expression levels of eight reference gene candidates (18S, TBP, HMBS, YWHAZ, ACTB, HPRT1, GAPDH and EEF1A2) in ten tissues types sourced from Boer goats. The optimal reference gene combination was selected according to the results determined by geNorm, NormFinder and Bestkeeper software packages. The analyses showed that tissue is an important variability factor in genes expression stability. When all tissues were considered, 18S, TBP and HMBS is the optimal reference combination for calibrating quantitative PCR analysis of gene expression from goat tissues. Dividing data set by tissues, ACTB was the most stable in stomach, small intestine and ovary, 18S in heart and spleen, HMBS in uterus and lung, TBP in liver, HPRT1 in kidney and GAPDH in muscle. Overall, this study provided valuable information about the goat reference genes that can be used in order to perform a proper normalisation when relative quantification by qRT-PCR studies is undertaken.

• Macromolecular Research
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• 제17권4호
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• pp.245-249
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• 2009

A fluorescing, copolymer(Q)-bearing, quaternary ammonium pendant was mixed with excess natural salmon sperm DNA with a molecular weight of $1.3{\times}10^6$(2,000 base pairs) to afford highly fluorescing, complex mixtures. The fluorescence life-time of the polymer Q was greatly increased when mixed with DNA: for the mixture of Q:DNA=1:750 the fast and slow decay lifetimes increased from ca. 10 to 100 ps and from 20 ps to ca. 1 ns, respectively. The enhanced fluorescence of the mixtures was ascribed to efficient compartmentalization and reduced conformational relaxation of the polymer Q by complexation with excess DNA.

• Journal of Ginseng Research
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• 제33권3호
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• pp.240-243
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• 2009

원료삼 생산에 있어 총수익(R)을 결정하는 4요인 승법 모형을 설정하고 (R=AYQP) 세 농가의 수납실적 수치를 사용하여 총수익 결정식임을 확인하였다. 4요소는 자본요소인 재배면적(A), 기술요소인 단위수량(Y)과 가중평균 품질등급(Q) 그리고 시장요소인 평균등급 가격(P)의 3요소로 해석하였다. 기술요소인 YQ는 자본(A)과 수익(R)의 직선모형에서 기울기가 되어 단위자본의 수익창출계수임을 밝혔다. 수익결정식은 인삼산업의 발전이 재배수량과 품질 향상기술 YQ를 증대시켜 면적(A)을 감소시킴으로서 수익(R)을 높여 시장가격(P)을 낮추는데 있음을 보여주었다.

• 한국환경농학회지
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• 제36권2호
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• pp.135-139
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• 2017

BACKGROUND: Identification and characterization of endophytic yeasts inhabiting the roots of Ulmus parvifolia Jacq. and Quercus salicina Blume require biotechnological and culture-based techniques. METHODS AND RESULTS: Homogenized U. parvifolia and Q. salicina root samples were spread onto four types of agar medium containing ancgtibiotics, L-sorbose, and Triton X-100. In total, 25 yeast strains were isolated and subjected to phylogenetic analysis based on their internal transcribed spacer region sequences. The results revealed that the yeast genera Cyberlindnera (12 isolates) and Cryptococcus (1 isolate) were associated with roots of U. parvifolia; and the genera Rhodotorula (8 isolates), Trichosporon (3 isolates), and Kluyveromyces (1 isolate) were associated with roots of Q. salicina. Additionally, a Kluyveromyces isolate produced a detectable level of bioethanol. The yeast strains reported herein may be used in industrial production of biosurfactants and bioethanol. CONCLUSION: Our findings revealed that the endophytic yeast genera Cyberlindnera and Cryptococcus predominated in roots of U. parvifolia; and the genera Rhodotorula (8 isolates), Trichosporon (3 isolates), and Kluyveromyces (1 isolate) predominated in roots of Q. salicina. Additionally, Kluyveromyces isolates produced a detectable level of bioethanol.

• Bulletin of the Korean Chemical Society
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• 제32권5호
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• pp.1604-1612
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• 2011

We developed three-dimensional quantitative structure activity relationship (3D-QASR) models for 17 adamantyl derivatives as P-glycoprotein (P-gp) inhibitors. Eighteen different partial charge calculation methods were tested to check the feasibility of the 3D-QSAR models. Best predictive comparative molecular field analysis (CoMFA) model was obtained with the Austin Model 1-Bond Charge Correction (AM1-BCC) atomic charge. The 3D-QSAR models were derived with CoMFA and comparative molecular similarity indices analysis (CoMSIA). The final CoMFA model ($q^2$ = 0.764, $r^2$ = 0.988) was calculated with an AM1-BCC charge and electrostatic parameter, whereas the CoMSIA model ($q^2$ = 0.655, $r^2$ = 0.964) was derived with an AM1-BCC charge and combined steric, electrostatic, hydrophobic and HB-acceptor parameters. Leave-five-out (LFO) cross-validation was also performed, which yielded good correlation coefficient for both CoMFA (0.801) and CoMSIA (0.656) models. Robustness of the developed models was checked further with 1000 run bootstrapping analyses, which gave an acceptable correlation coefficient for CoMFA (BS-$r^2$ = 0.997, BS-SD = 0.003) and CoMSIA (BS-$r^2$ = 0.996, BS-SD = 0.018).

• 한국환경농학회지
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• 제30권4호
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• pp.367-376
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• 2011

논 토양의 미생물 생태 다양성을 조사하기 위한 효과적인 방법으로 qRT-PCR을 적용하고자 본 연구를 수행하였다. 논 토양 미생물의 gDNA를 분리하기 위하여 Mo Bio kit를 사용한 효과적이고 안정적인 gDNA 분리 방법을 확립하였다. 논 토양 미생물 다양성을 qRT-PCR로 검출하기 위하여 bacteria를 세분한 ${\alpha}$-Proteobacteria, ${\beta}$-Proteobacteria, Actinobacteria, Bacteroidetes, Firmicutes 다섯 가지 문과 전체 bacteria, 전체 fungi를 구분할 수 있는 특이 primer set을 선정하여 다양한 조건의 시험을 통하여 최종 조건을 확립하였으며 재현성 실험을 통하여 방법의 유의성을 검증하였다.

• Journal of Forest and Environmental Science
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• 제27권3호
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• pp.143-149
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• 2011

The possibility of employing biomass of Salix viminalis cv. Q683 as a resource of bio-energy was evaluated. The chemical analysis of S. viminalis cv. Q683 leaf biomass showed components such as, extractives (2.57%), lignin (39.06%), hemicellulose (21.61%), and cellulose (37.83%), whereas, its stem was composed of extractives (1.67%), lignin (23.54%), hemicellulose (33.64%), and cellulose (42.03%). The biomass of S. viminalis cv. Q683 was saccharified using two enzymes celluclast and viscozyme. The saccharification of S. viminalis cv. Q683 biomass was influenced by enzymes and their strengths. The optimal enzyme combination was found to be celluclast (59 FPU/g substrate) and viscozyme (24 FBG/g substrate). On saccharification the glucose from leaf and stem biomass was 7.5g/L and 11.7g/L, respectively after 72 hr of enzyme treatment. The biomass and enzyme-treated biomass served as the feedstock for ethanol production by fermentation. The ethanol production from stem and leaf biomass was 5.8 g/L and 2.2 g/L respectively, while the fermentation of the enzymatic hydrolysates yielded 5 g/L to 8 g/L bioethanol in 72 hours.