• Title/Summary/Keyword: C5 Protein

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Sasa borealis leaves extract improves insulin resistance by modulating inflammatory cytokine secretion in high fat diet-induced obese C57/BL6J mice

  • Yang, Jung-Hwa;Lim, Hyeon-Sook;Heo, Young-Ran
    • Nutrition Research and Practice
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    • v.4 no.2
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    • pp.99-105
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    • 2010
  • Obesity is considered a mild inflammatory state, and the secretion of inflammation-related cytokines rises as adipose tissue expands. Inflammatory cytokines, including tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), interlukin 6 (IL-6) and monocyte-chemoattractant protein 1 (MCP-1), are modulated by adipose tissue and known to play an important role in insulin resistance which is the common characteristics of obesity related disorders. In this study we analyzed the effects of Sasa borealis leaves extract on inflammatory cytokines and insulin resistance in diet induced obese C57/BL6J mice. The obese state was induced by a high fat diet for 20 weeks and then the mice were divided into two groups; obese control group (OBC, n = 7) and experimental group (OB-SBE, n = 7). The OBC group was fed a high fat diet and the OB-SBE group was fed a high fat diet containing 5% Sasa borealis leaves extract (SBE) for 12 weeks. We also used mice fed a standard diet as a normal control (NC, n = 7). The body weight and adipose tissue weight in the OB group were significantly higher than those in the NC group. The effects of the high fat diet were reduced by SBE treatments, and the body weight and adipose tissue deposition in the OB-SBE group were significantly decreased compared to the OBC group. The OBC group showed higher serum glucose and insulin levels which resulted in a significant increase of incremental area under the curve (IAUC) and HOMA-IR than the NC group. Also, serum leptin, TNF-${\alpha}$, and IL-6 levels were significantly higher in the OBC group than in the NC group. In contrast, the OB-SBE group showed a reversal in the metabolic defects, including a decrease in glucose, insulin, IAUC, HOMA-IR, TNF-${\alpha}$, IL-6 and leptin levels. These results suggest that BSE can suppress increased weight gain and/or fat deposition induced by a high fat diet and theses effects are accompanied by modulation of the inflammatory cytokines, TNF-${\alpha}$ and IL-6 secretion resulting in improved insulin resistance.

Pharmacokinetics of Recombinant Human Granulocyte Colony Stimulating Factor (rhG-CSF) Following Intravenous, Intramuscular and Subcutaneous Administration of HM10411 and Filgrastim to Rats and Mice (인과립구 콜로니 자극인자 제제인 HM10411와 필그라스팀의 정맥, 근육 및 피하 주사시 흰쥐와 마우스에서의 약물 동태)

  • Kim, In-Wha;Lee, Sang-Hoon;Kim, Young-Min;Jung, Sung-Youb;Kwon, Se-Chang;Lee, Gwan-Sun;Chung, Suk-Jae;Shim, Chang-Koo
    • Journal of Pharmaceutical Investigation
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    • v.31 no.2
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    • pp.89-94
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    • 2001
  • The pharmacokinetics of recombinant human granulocyte colony stimulating factor (rhG-CSF) following intravenous (i.v.), intramuscular (i.m.) and subcutaneous (s.c.) administration of HM1041l-lyo and HM10411-liq (lyophilized and liquid formulations of rhG-CSF, recently under development by Hanmi Pharmaceutical Company) were studied in rats, and compared with that of Filgrastim (conventional formulation of rhG-CSF on market). The plasma concentration of rhG-CSF was quantified using a specific ELISA. The pharmacokinetic parameters of rhG-CSF, after i.v., i.m. and s.c. administration of Filgrastim, HM1041l-lyo and HM1041l-liq to rats at a rhG-CSF dose of $10\;{\mu}g/kg$, were almost identical among the three formulations. No dose-dependency was observed in the pharmacokinetic parameters of rhG-CSF following i.v. administration in the dose range of $5{\sim}100\;{\mu}g/kg$. rhG-CSF, after i.v. administration of the three preparations at a dose of $10\;{\mu}g/kg$ to rats, was detected at low levels in all of the body tissues with highest tissue/plasma ratio of $0.46{\sim}0.51$ for the kidney at 30 min after the administration. The pharmacokinetics of rhG-CSF, after i.v. administration to mice at a dose of $10\;{\mu}g/kg$, were comparable among the three formulations. In conclusion, HM10411-lyo and HM10411-liq exhibited similar pharmacokinetics for rhG-CSF with Filgrastim regandless of animal species. Considering the fact that HM10411 series, contrary to Filgrastim, are proteins lacking a methionine residue, the methionine moiety in rhG-CSF molecule does not appear to influence the pharmacokinetics of the protein significantly.

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버섯의 항암효과 -영자 균사체 다당류의 항종양 활성-

  • 이준우
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 1997.06b
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    • pp.13-14
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    • 1997
  • 영지의 약리효과는 triterpenoid계의 저분자와 polysaccharide인 고분자로 대별할 수 있다. 저분자 물질은 항염증작용, 항알러지작용, 간암세포 성장 억제작용, 혈압 강하작용, 혈소판 응집저해작용 등이 있으며, 고분자 물질은 혈압강하작용, 정혈작용, 고지혈증 개선작용, 혈당 강하작용, 면역 증강작용 및 항종양작용 등이 있는 것으로 알려졌다. 현재까지의 영지에 관한 연구는 주로 자실체에 대한 연구가 주류를 이루고 있으며 이들의 산지, 환경조건, 채집시기 및 실험자들에 따라 약리 효과 등에 많은 차이가 있다. 이들에 대한 효과가 영지의 대표적 약리효과라고 언급하기엔 부족한 점이 많으므로 약리효과가 우수하고 일관성이 있는 영지 유래 다당류를 얻기 위한 방법으로서는 균사체의 대량배양법 및 분리방법이 확립 되어야만 한다. 본 연구에서는 새로운 항암활성 다당류의 개발을 위해 국내 자생 영지를 채집하여 검색 및 계통적 분류를 행하였으며 분리된 균주의 최적 액체배양 조건, 다당류 분리 조건, 다당류의 물리화학적 특성 및 약리활성 등에 관한 연구를 실시하였다. 그 중 약리활성이 우수한 G- lucidum IY009 균주를 선발 하였다. 구조적 특징과 종양 면역 활성과의 관계 규명을 위해, G- lucidum IY009 배양균사체로부터 다당류를 T-AS, U-AS, M-AS, S 및 H로 분획 하였다. 추출조건에 따른 다당류 GLG의 수율은 열수 추출 분획이 2.98g/l로 가장 높았으며, U-AS및 T-AS에서 각각 2.32g/l, 2.07g/l이었다. GLG의 특성을 조사하기 위하여 수용성과 비수용성으로 분리한 결과 수용성 분획은 비수용성 분획보다 높은 당 함량을 나타냈으며, T-AS는 70.3%의 당과 7.8%의 단백질로 구성 되었다. GLG 대부분의 분획들은 60~93%의 glucose로 구성된 다당류 이었으며, 주로 $\beta$-glucose로 구성된 다당류 이었다. 아미노산은 Asp 및 Glu의 산성 아미노산과 Ala, Leu 등의 함량이 높게 나타났으며, 비알칼리 추출물에서 Ser과 Thr의 함량이 높게 나타났다. 다당류 T-AS는 평균 분자량이 2,000 kD와 12kD에서 주 peak를 나타냈으며, 수용성 분획의 평균 분자량은 12kD이고 비수용성 분획은 36~2,000 kD의 평균 분자량 분포를 갖는 것으로 나타났다. IR과 NMR 분석 결과 890 cm-1에서 흡수 peak를 나타내어 $\beta$-(1,3)0glucan과 $\beta$-(1,6)-glucan의 구조를 갖는 다당류로 확인 되었다. T-AS 분획은 C:H:O:N의 함량비가 38.9:5.7:49.6:1.84%이며, 이 물질의 융점은 163 $^{\circ}C$로 연한 갈색을 나타낸다. 분리된 GLG의 항암활성 기전 규명을 위해, in vivo 항암실험, 항보체 활성능, 항체 생성능, serum protein 분비능, 대식세포의 탐식능과 활성능 및 세포간 물질 분비 등의 상관관계를 조사하였다. 다당류 GLG 분획물들 가운데 항보체의 활성이 높았던 분획은 sarcome 180에 대한 항암 활성이 높게 나타났다. 다당류 T-AS의 보체 활성화 기작은 classical과 alternative complement pathway의 양 경로를 통해 활성화 되었다. T-AS 분획은 mouse내의 특정 혈청단백을 증가시켰으며, 항체 생성능의 증가가 관찰되어 effect T 세포의 활성화가 나타나고 있음을 알 수 있었다. T-AS는 생체내 투여시에 대식세포의 탐식능이 증진되었으며, 대식세포 기능 저해제에 의한 대식세포의 기능 저해 현상이 회복되었다. 이와 같은 결과들로부터, T-AS의 항암 활성은 활성화된 보체 성분 및 당 수용체들이 존재하는 대식세포의 개입을 시사한다.

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유산균의 항암효과

  • 배형석
    • Proceedings of the Korean Journal of Food and Nutrition Conference
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    • 1997.06b
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    • pp.15-17
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    • 1997
  • 암 발생의 원인 중 80% 이상이 식사습관과 환경오염에 있는 것으로 인정되고 있다. 음식물, 담배, 술, 대기오염, 스트레스, 자외선이 그 대표적인 원인물질로 꼽을 수 있으며 그 중에서도 매일 섭취하는 음식물이 가장 중요한 발암요인으로 지적되고 있다. 대장암과 유방암의 발생에 대한 Wynder 등의 역학조사에서도 식사습관이 암 발생에 중요함을 시사 하고 있다. 동물성 단백질과 지방의 다량 섭취가 대장암과 유방암의 발생을 증가 시키고 섬유질이 풍부한 곡류와 야채의 섭취는 대장암 발생을 억제한다는 상관관계가 밝혀졌다. 그러나 우리가 늘 섭취하는 음식물 자체는 대장암과 유방암을 유발하는 기능이 거의 없다. 섭취된 음식물이 암을 일으키려면 장내 부패 미생물의 분해작용에 의하여 발암물질로 변환되는 과정이 필요하다. 그 발암물질들이 장관으로 흡수 자극함으로써 암을 유발할 수 있다. 반대로 일부장내 미생물들은 장내 발암물질들을 무독화 하거나 숙주의 면역기능을 증강 시킴으로써 암 발생을 억제할 수도 있다는 사실을 간과해서는 안 될 것이다. Mitsuoka는 장내 미생물이 암을 유발하는 중요한 요인이라고 강조하였다. Veer 등은 유산균 발효유를 많이 섭취하면 유방암 발생이 억제됨을, 국제암연구위원회는 섬유질을 많이 섭취하고 있는 핀란드 쿠피오의 거주자들에게는 덴마크의 코펜하겐에 거주하는 사람들에 비하여 대장암 발생율이 1/4에 불과하고 분변내 유산간균수는 100배 높은 사실을 역학조사에서 밝혔다. 이 외에 유산균과 발효유제품의 항암효과에 대한 실험결과들이 많이 발표되었다. 여기에서는 유산균의 항암효과에 대한 지금까지의 관련 자료들을 요약, 정리하여 고찰하고자 한다.높은 당 함량을 나타냈으며, T-AS는 70.3%의 당과 7.8%의 단백질로 구성 되었다. GLG 대부분의 분획들은 60~93%의 glucose로 구성된 다당류 이었으며, 주로 $\beta$-glucose로 구성된 다당류 이었다. 아미노산은 Asp 및 Glu의 산성 아미노산과 Ala, Leu 등의 함량이 높게 나타났으며, 비알칼리 추출물에서 Ser과 Thr의 함량이 높게 나타났다. 다당류 T-AS는 평균 분자량이 2,000 kD와 12kD에서 주 peak를 나타냈으며, 수용성 분획의 평균 분자량은 12kD이고 비수용성 분획은 36~2,000 kD의 평균 분자량 분포를 갖는 것으로 나타났다. IR과 NMR 분석 결과 890 cm-1에서 흡수 peak를 나타내어 $\beta$-(1,3)0glucan과 $\beta$-(1,6)-glucan의 구조를 갖는 다당류로 확인 되었다. T-AS 분획은 C:H:O:N의 함량비가 38.9:5.7:49.6:1.84%이며, 이 물질의 융점은 163 $^{\circ}C$로 연한 갈색을 나타낸다. 분리된 GLG의 항암활성 기전 규명을 위해, in vivo 항암실험, 항보체 활성능, 항체 생성능, serum protein 분비능, 대식세포의 탐식능과 활성능 및 세포간 물질 분비 등의 상관관계를 조사하였다. 다당류 GLG 분획물들 가운데 항보체의 활성이 높았던 분획은 sarcome 180에 대한 항암 활성이 높게 나타났다. 다당류 T-AS의 보체 활성화 기작은 classical과 alternative complement pathway의 양 경로를 통해 활성화 되었다.

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Fabrication of a Partial Genome Microarray of the Methylotrophic Yeast Hansenula polymorpha: Optimization and Evaluation of Transcript Profiling

  • OH , KWAN-SEOK;KWON, OH-SUK;OH, YUN-WI;SOHN, MIN-JEONG;JUNG, SOON-GEE;KIM, YONG-KYUNG;KIM, MIN-GON;RHEE, SANG-KI;GERD GELLISSEN,;KANG, HYUN-AH
    • Journal of Microbiology and Biotechnology
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    • v.14 no.6
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    • pp.1239-1248
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    • 2004
  • The methylotrophic yeast Hansenula polymorpha has been extensively studied as a model organism for methanol metabolism and peroxisome biogenesis. Recently, this yeast has also attracted attention as a promising host organism for recombinant protein production. Here, we describe the fabrication and evaluation of a DNA chip spotted with 382 open reading frames (ORFs) of H. polymorpha. Each ORF was PCR-amplified using gene-specific primer sets, of which the forward primers had 5'-aminolink. The PCR products were printed in duplicate onto the aldehyde-coated slide glasses to link only the coding strands to the surface of the slide via covalent coupling between amine and aldehyde groups. With the partial genome DNA chip, we compared efficiency of direct and indirect cDNA target labeling methods, and found that the indirect method, using fluorescent-labeled dendrimers, generated a higher hybridization signal-to-noise ratio than the direct method, using cDNA targets labeled by incorporation of fluorescence-labeled nucIeotides during reverse transcription. In addition, to assess the quality of this DNA chip, we analyzed the expression profiles of H. polymorpha cells grown on different carbon sources, such as glucose and methanol, and also those of cells treated with the superoxide­generating drug, menadione. The profiles obtained showed a high-level induction of a set of ORFs involved in methanol metabolism and oxidative stress response in the presence of methanol and menadione, respectively. The results demonstrate the sensitivity and reliability of our arrays to analyze global gene expression changes of H. polymorpha under defined environmental conditions.

Heterologous Expression and Characterization of Glycogen Branching Enzyme from Synechocystis sp. PCC6803

  • Lee, Byung-Hoo;Yoo, Young-Hee;Ryu, Je-Hoon;Kim, Tae-Jip;Yoo, Sang-Ho
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1386-1392
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    • 2008
  • A gene (sll0158) putatively encoding a glycogen branching enzyme (GBE, E.C. 2.4.1.18) was cloned from Synechocystis sp. PCC6803, and the recombinant protein expressed and characterized. The PCR-amplified putative GBE gene was ligated into a pET-21a plasmid vector harboring a T7 promoter, and the recombinant DNA transformed into a host cell, E. coli BL21(DE3). The IPTG-induced enzymes were then extracted and purified using Ni-NTA affinity chromatography. The putative GBE gene was found to be composed of 2,310 nucleotides and encoded 770 amino acids, corresponding to approx. 90.7 kDa, as confirmed by SDS-PAGE and MALDI-TOF-MS analyses. The optimal conditions for GBE activity were investigated by measuring the absorbance change in iodine affinity, and shown to be pH 8.0 and $30^{\circ}C$ in a 50 mM glycine-NaOH buffer. The action pattern of the GBE on amylose, an $\alpha$-(1,4)-linked linear glucan, was analyzed using high-performance anion-exchange chromatography (HPAEC) after isoamylolysis. As a result, the GBE displayed $\alpha$-glucosyl transferring activity by cleaving the $\alpha$-(1,4)-linkages and transferring the cleaved maltoglycosyl moiety to form new $\alpha$-(1,6)-branch linkages. A time-course study of the GBE reaction was carried out with biosynthetic amylose (BSAM; $M_p{\cong}$8,000), and the changes in the branch-chain length distribution were evaluated. When increasing the reaction time up to 48 h, the weight- and number-average DP ($DP_w$ and $DP_n$) decreased from 19.6 to 8.7 and from 17.6 to 7.8, respectively. The molecular size ($M_p$, peak $M_w{\cong}2.45-2.75{\times}10^5$) of the GBE-reacted product from BSAM reached the size of amylose (AM) in botanical starch, yet the product was highly soluble and stable in water, unlike AM molecules. Thus, GBE-generated products can provide new food and non-food applications, owing to their unique physical properties.

Assessment of Relationship between Fyn-related Kinase Gene Polymorphisms and Overweight/Obesity in Korean Population

  • Jung, Mi-Young;Kim, Bum-Shik;Kim, Youn-Jung;Koh, In-Song;Chung, Joo-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.12 no.2
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    • pp.83-87
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    • 2008
  • The fyn-related kinase (FRK) belongs to the tyrosine kinase family of protein kinases. Recent studies have shown that Frk affects pancreatic beta cell number during embryogenesis and promotes beta cell cytotoxic signals in response to streptozotocin. To investigate the genetic association between FRK polymorphisms and the risk of obesity in Korean population, single nucleotide polymorphisms (SNPs) in the FRK gene region were selected and analyzed. The body mass index (BMI) was calculated, and biochemical data (systolic blood pressure, diastolic blood pressure, hemoglobin A1C, triglyceride, total cholesterol, high density lipoprotein, and low density lipoprotein) of blood sample from each subject were also measured. One hundred fifty five healthy control and 204 overweight/obesity subjects were recruited. Genotype frequencies of six SNPs [rs6568920 (+8391G>A), rs3756772 (+56780A>G), rs3798234 (+75687C>T), rs9384970 (+68506G>A), rs1933739 (+72978G>A), and rs9400883 (+75809A>G)] in the FRK gene were determined by Affymetrix Targeted Genotyping Chip data. According to the classification of Korean Society for the Study of Obesity, control (BMI 18 to < 23) and overweight/obesity (BMI$\geq$23) subjects were recruited. For the analysis of genetic data, EM algorithm, SNPStats, Haploview, HapAnalyzer, SNPAnalyzer, and Helixtree programs were used. Multiple logistic regression analysis (codominant, dominant, and recessive models) was performed. Age and gender as covariates were adjusted. For biochemical data, Student's t test was used. The mean value of BMI in the control and overweigh/obesity groups was 21.1${\pm}$1.2 (mean${\pm}$SD) and 25.6${\pm}$2.0, respectively. All biochemical data of the overweight/obesity group were statistically significance, compared with the control group. Among six SNPs, two linkage disequilibrium (LD) blocks were discovered. One block consisted of rs1933739 and rs9400883, and the other comprised rs3756772 and rs3798234. One SNP (rs9384970, +68506G>A) showed an association with overweight/obesity in the codominant model (p=0.03). Interestingly, the AA genotype distribution in the overweight/obesity group (n=7, 3.5%) was higher than those in the control group (n=1, 0.6%), which is not found in either Japanese or Chinese subjects. Therefore, the AA genotype of rs9384970 may be a risk factor for development of obesity in Korean population. The results suggest that FRK may be associated with overweight/obesity in Korean population.

Effect of Saponin and Non-saponin of Panax Ginseng on the Blood Pressure in the Renovascular Hypertensive Rats (신성고혈압백서에서 혈압에 미치는 고려홍삼사포닌과 비사포닌의 효과)

  • Jeon Byeong Hwa;Kim Hoe Suk;Chang Seok Jong
    • Journal of Ginseng Research
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    • v.23 no.2 s.54
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    • pp.81-87
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    • 1999
  • The effect of saponin and non-saponin of Panax Red Ginseng on the blood pressure and nitric oxide production were investigated in the conscious free moving one-kidney, one-clip Goldbaltt hypertensive (lK, 1C-GBH) rats. Mean blood pressure in the control and lK, 1C-GBH rats was decreased by the administration of ginseng saponin (100 mg/kg, i.v.). The hypotensive effect induced by ginseng saponin was reached maximum at 2-4 minutes and was slowly recovered to the initial level of blood pressure. Also ginseng saponin induced reflex tachycardia in the conscious both rats. Contrast to the response induced by ginseng saponin, hypotensive effect induced by non-saponin of panax ginseng is minimal. Plasma nitric oxide concentration was increased by the treatment of ginseng saponin (100 mg/kg, i.p for 5 days) in both rats. It has been shown by western blotting that the expression level of the protein for endothelial nitric oxide synthase (eNOS) in the aorta of rats was not increased by the treatment of ginseng saponin (100 mg/kg, i.p). However, eNOS activity in aortic homogenates of both rats were increased by the treatment of ginseng saponins. From the above results, the hypotensive effect of saponin was greater than that of non-saponin of Panax Red Ginseng. The lowering effect of blood pressure by ginseng saponin may be due to the increase of plasma nitric oxide concentration via the increase of endothelial nitric oxide synthase activity in the renovascular hypertensive and control rats.

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Insect Resistance of Tobacco Plant Expressing CpBV-ELP1 Derived from a Polydnavirus (폴리드나바이러스 유래 CpBV-ELP1 발현 담배의 내충성)

  • Kim, Eunseong;Kim, Yonggyun
    • Korean journal of applied entomology
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    • v.56 no.1
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    • pp.19-28
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    • 2017
  • Polydnaviruses (PDVs) are a group of double-stranded DNA viruses symbiotic to some endoparasitoid wasps. Cotesia plutellae bracovirus (CpBV) is a PDV symbiotic to an endoparasitoid wasp, C. plutellae, parasitizing young larvae of Plutella xylostella. An early expressed gene, CpBV-ELP1, plays an important role in the parasitism by suppressing host cellular immunity by its cytotoxic activity against hemocytes. This study aimed to test its oral toxicity against insect pest by expressing it in a recombinant tobacco plant. A recombinant CpBV-ELP1 protein was produced using a baculovirus expression system and secreted to cell culture medium. The cell cultured media were used to purify CpBV-ELP1 by a sequential array of purification steps: ammonium sulfate fractionation, size exclusion chromatography, and ion exchange chromatography. Purified rCpBV-ELP1 exhibited a significant cytotoxicity against Spodoptera exigua hemocytes. CpBV-ELP1 was highly toxic to the fifth instar larvae of S. exigua by injection to hemocoel. It also showed a significant oral toxicity to fifth instar larvae of S. exigua by a leaf-dipping assay. CpBV-ELP1 was cloned into pBI121 vector under CaMV 35S promoter with opaline synthase terminator. Resulting recombinant vector (pBI121-ELP1) was used to transform Agrobacterium tumefaciens LBA4404. The recombinant bacteria were then used to induce callus of a tobacco (Nicotiana tabacum Xanthi) leaves and subsequent generation (T1) plants were selected. T1 generation tobacco plants expressing CpBV-ELP1 gave significant insecticidal activities against S. exigua larvae. These results suggest that CpBV-ELP1 gene can be used to control insect pests by constructing transgenic crops.

Studies on the Control of Summer Depression of Pasture Plants -Effect of Mulching on the Productivity and Chemical Composition of Pasture Plants- (목초(牧草)의 하고성(夏枯性) 방제(防除)에 관(關)한 연구(硏究) -초지피복(草地被覆)이 목초(牧草) 생산성(生産性) 및 영양성분(營養成分)에 미치는 영향(影響)-)

  • Kim, Jong-Woo
    • Korean Journal of Agricultural Science
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    • v.3 no.2
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    • pp.184-191
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    • 1976
  • This experiment was conducted for the control of summer depression of cool-season pasture plant. Orchard grass, tall fescue, red clover, alfalfa, and crabgrass were mulched by barley straw with depths of $100g/m^2$, $200g/m^2$ and $300g/m^2$ for 45 days -from July 1, to August 14-, and the effects on growth, yield and chemical composition were observed. The results obetain were as follow: Temperature decrease of $1.0{\sim}3.04^{\circ}C$ was followed after barley straw mulching on the grassland as compared with the control. The growth of the grasses after mulching was hastened, i. e. plant height was increasd 6.0~45% as compared with the control. Barley straw mulching with $100g/m^2$ supported the increase in yield of the grasses with an average of 32.77%, however, average yield was decreased by 7.75% with $300g/m^2$ mulching, apparently due to the rottening of grasses. Chemical compositions of mulched grasses were varied; contents of crude protein and nitrogen free extract were higher, but crude fats, fibre, and ashes were lower than those of the control.

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